Tissue filler compositions and methods of use

ABSTRACT

The present disclosure relates to methods for stimulating collagen synthesis, cosmetic enhancement of soft tissue and/or inhibiting or treating scarring comprising administering a composition to an area to be treated within a soft tissue, wherein the composition comprises a tissue filler medium and a fluorophore; and illuminating the area with light having a wavelength which can be absorbed by the fluorophore.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation application of U.S. patentapplication Ser. No. 15/301,189, filed on Sep. 30, 2016, which is anational stage application under 35 U.S. C. § 371 of InternationalApplication No. PCT/CA2015/050261, filed Apr. 1, 2015, which claims thebenefit of an priority to U.S. provisional patent application No.61/973,659; filed Apr. 1, 2014, the content of which is hereinincorporated in its entirety by reference. International ApplicationPCT/CA2015/059126 was published under PCT Article 21(2) in English.

FIELD OF TECHNOLOGY

The present disclosure generally relates to tissue filler compositionsand methods of use. Specifically, but not exclusively, the presentdisclosure relates to tissue filler compositions which can be injectedor implanted into soft tissues.

BACKGROUND OF THE DISCLOSURE

Tissue fillers are used in the medical and cosmetic fields for tissueaugmentation, regeneration or re-shaping. In the cosmetic field, tissuefillers are known as dermal fillers and are injectable materials whichcan improve the appearance of skin by filling of wrinkles and folds,replacing lost tissue, or by facial contouring such as re-shaping thecheeks, chin or jaw. Tissue fillers are also used for treating scars bytemporarily elevating a depressed scar.

It is an object of the present disclosure to provide new and improvedtissue filler compositions and methods useful in medical and cosmeticfields.

SUMMARY OF SOME EMBODIMENTS OF THE PRESENT DISCLOSURE

According to various aspects, the present disclosure providescompositions and methods useful as tissue filler for medical andcosmetic indications.

According to various aspects, the present disclosure relates to a methodfor stimulating collagen synthesis comprising: administering acomposition to an area to be treated within a soft tissue, wherein thecomposition comprises a tissue filler medium and a fluorophore;illuminating the area with light having a wavelength which can beabsorbed by the fluorophore; wherein the method stimulates collagensynthesis in the area.

According to various aspects, the present disclosure relates to a methodfor cosmetic enhancement of soft tissue comprising: intradermally orsubdermally administering a composition to an area to be treated,wherein the composition comprises a tissue filler medium and afluorophore; illuminating the area with light having a wavelength whichcan be absorbed by the fluorophore; wherein the method stimulatescollagen synthesis in the area to cosmetically enhance the soft tissue.

According to various aspects, the present disclosure relates to a methodfor skin rejuvenation comprising: intradermally or subdermallyadministering a composition to an area to be treated, wherein thecomposition comprises a tissue filler medium and a fluorophore;illuminating the area with light having a wavelength which can beabsorbed by the fluorophore; wherein the method stimulates collagensynthesis in the area to cosmetically rejuvenate the skin.

According to various aspects, the present disclosure relates to a methodfor inhibiting or treating scarring, the method comprising:administering a composition to an area to be treated within or around ascar or a wound, wherein the composition comprises a tissue fillermedium and a fluorophore; illuminating the area with light having awavelength which can be absorbed by the fluorophore; wherein the methodstimulates collagen synthesis in the area to prevent or reduce scarformation.

According to various aspects, the present disclosure relates to a methodfor promoting wound healing, the method comprising: administering acomposition to an area to be treated within a wound, wherein thecomposition comprises a tissue filler medium and a fluorophore;illuminating the area with light having a wavelength which can beabsorbed by the fluorophore; wherein the method stimulates wound healingin the area.

By means of certain embodiments of the present disclosure, light emittedby the fluorophore can induce or stimulate collagen synthesis in thesoft tissue around the composition where there otherwise would be nocollagen synthesis or where the collagen synthesis would be minimal. Forexample, the collagen synthesis induced by the fluorophore may be inaddition to any collagen synthesis inherently induced by the tissuefiller medium by mechanical means (e.g. when the tissue filler mediumcomprises hyaluronic acid) or by a foreign body response (e.g. when thetissue filler medium is a nonbiodegradable material). When thefluorophore is activated to emit fluorescent or phosphorescent light,the surrounding soft tissues are illuminated with a broader spectrum oflight than if illuminated from outside the dermis or from a single lightsource. Also, the surrounding soft tissues may be illuminated withshorter wavelengths of light than those able to reach those soft tissuestransdermally. In this way, a more efficient and wavelength specificillumination of the soft tissues, such as dermal/epidermal/subdermallayers can be achieved which may lead to a therapeutic or cosmeticeffect.

In certain embodiments, the area to be treated is soft tissue. The areato be treated may be on any part of the body such as on a face, neck,ears, breasts, buttocks, arms, armpits, hands, genitalia, legs or feetof a human subject. In certain embodiments, the area to be treated is inor around a scar. The scar may be a post-surgical scar. The scar can bean old or a fresh scar. In certain embodiments, the area to be treatedis in or around a wound. The wound may be an acute or a chronic wound,including burns. In certain embodiments, the area to be treated includesstretch marks. The area to be treated may be in or around a stretchmark.

In certain embodiments, the administering of the composition is byinjection. The injection can be performed using a needle. The needle canhave a gauge of 27 G to 40 G, typically 30 G or 32 G. The administeringby injection can be a continuous injection, known as linear threading,or serial punctures to deposit microdroplets. In certain otherembodiments, the administering of the composition is by implantation.

In certain embodiments, the composition is a cohesive gel. Thecomposition may be a hydrated gel. The composition may be transparent ortranslucent. In certain embodiments, the tissue filler medium is acohesive gel. The tissue filler medium may be a hydrated gel. The tissuefiller medium may be transparent or translucent.

In certain embodiments, the tissue filler medium retains the fluorophorewithin the composition during administering of the composition, and atleast during a portion of the illumination. Alternatively, in certainembodiments, the tissue filler medium/composition may allow thefluorophore to leach from the composition/tissue filler medium afteradministering to the tissue.

In certain embodiments, the tissue filler medium is biodegradable. Thetissue filler medium may comprise any biodegradable and biocompatiblematerial such as hyaluronic acid (HA), collagen, poly-L-lactic acid.Complete biodegradation may occur in about 3-18 months. The fluorophoremay or may not affect the biodegradation rate of the tissue fillermedium.

In certain embodiments, the tissue filler medium comprises cross-linkedhyaluronic acid (cross-linked HA). The cross-linked hyaluronic acid maybe about 0.1% to about 2%, about 2% to about 30%, about 2% to about 25%,about 2% to about 20%, about 2% to about 15%, about 2% to about 10%,about 2% to about 5%, 4% to about 30%, about 4% to about 25%, about 4%to about 20%, about 4% to about 15%, about 4% to about 10%, about 4% toabout 5%, of the composition. In certain embodiments, the cross-linkedhyaluronic acid is in particulate form. The particles may be hydrated.The particles may be cohesive. In certain embodiments, the fluorophoreis within the cross-linked hyaluronic acid particle.

In certain embodiments, the composition further comprises an injectablemedium supporting the particles. The injectable medium may be a fluid.The injectable medium may be less viscous and/or less cohesive than theparticles. For example, the particles may be cohesive and the injectablemedium non-cohesive. The fluorophore may be in the injectable medium orthe particles, or both. In certain embodiments, the injectable mediumcomprises hyaluronic acid which is non-cross linked or relatively lesscross-linked than the cross-linked hyaluronic acid particles.

In certain embodiments, the composition further comprises lightreflecting particles. The light reflecting particles may be a glass orsilicon dioxide.

In certain embodiments, the fluorophore is a hydrophilic chromophore.The fluorophore may be water soluble. In certain embodiments, thefluorophore is not in a liposomal form in the composition. In certainembodiments, the fluorophore can be activated by light having awavelength in the visible range. In certain embodiments, the fluorophoredoes not absorb light in the UV range of the electromagnetic spectrum.In certain embodiments, when activated, the fluorophore can emit lighthaving a wavelength in the visible range. The emitted light can bewithin one or more of the violet, blue, green, yellow, orange or redportions of the electromagnetic spectrum. Alternatively, the fluorophorecan emit in the infrared range. In certain embodiments, the fluorophorecan be photobleached after illumination of the area. In certainembodiments, the fluorophore may be photobleached after 5, 10, 15, 20,25, 30, 35, 40, 45, 50, 55, 60 or 65 minutes of illumination. Theillumination may be continuous or pulsed. The illumination may comprisere-illumination of the composition or the area after a few hours ordays. In certain embodiments, by means of the photobleaching,photosensitivity is reduced or avoided. In certain embodiments, thefluorophore is not a photosensitizing agent requiring to be metabolized.For example, in certain embodiments, the fluorophore is not porphyrin,porphyrinogen, hematoporphyrin, pheophorbide, chlorin, bacteriochlorin,iso-bacteriochlorin and dihydro- and tetrahydro-tetrapyrroles. Incertain embodiments, the fluorophore does not bind to native cells orcellular structures. In certain embodiments, the area is illuminated atthe same time as or immediately after intradermal or subdermaladministration. In other words, there is no incubation period betweenadministering of the composition and illumination.

In certain embodiments, the illumination is for a time sufficient toactivate the fluorophore. In certain embodiments, the illumination isfor a time sufficient to photobleach the fluorophore. In certainembodiments, the illumination is for 1-30 seconds, 15-45 seconds, 30-60seconds, 0.75-1.5 minutes, 1-2 minutes, 1.5-2.5 minutes, 2-3 minutes,2.5-3.5 minutes, 3-4 minutes, 3.5-4.5 minutes, 4-5 minutes, 5-10minutes, 10-15 minutes, 15-20 minutes, 20-25 minutes, or 25-30 minutes.The treatment time may range up to about 90 minutes, about 80 minutes,about 70 minutes, about 60 minutes, about 50 minutes, about 40 minutesor about 30 minutes.

In certain embodiments, the composition is illuminated by a light sourcepositioned external to the dermis. In other words, illumination istransdermally.

In certain embodiments, intradermally or subdermally administrationprovides a track of HA-stimulated cells from the external surface to thedermis, thereby creating a light duct from the light source to theintradermal or subdermal in order to activate the composition.

In certain embodiments, the method further comprises applying a topicalcomposition over the treatment area, wherein the topical compositioncomprises a fluorophore. The fluorescent compound may have an emissionspectrum which can activate the fluorophore in the intradermal/subdermalcomposition. In this way a deeper illumination of soft tissues may beobtained. In certain embodiments, the topical composition is appliedprior to light illumination.

According to various aspects, the present disclosure relates to a methodfor skin rejuvenation comprising: intradermally or subdermallyadministering a composition to an area to be treated, wherein thecomposition comprises a tissue filler medium and a fluorophore;topically applying a biophotonic composition to the skin above the areato be treated; and illuminating the topical biophotonic composition withlight having a wavelength which can be absorbed by a chromophore in thetopical biophotonic composition to cause the chromophore to emit light;wherein the fluorophore can absorb the emitted light from the topicalbiophotonic composition, wherein the method stimulates collagensynthesis in the area to cosmetically rejuvenate the skin. In certainembodiments, the composition is injectable and can be injected through a23-40 gauge needle. In certain embodiments, the composition is injectedunder a wrinkle or a fold or a scar to provide an immediate lift to thedefect. Illumination of the composition may then stimulate collagensynthesis around the injected composition to provide further skinrejuvenation.

From another aspect there is provided a tissue filler compositioncomprising: a tissue filler medium; and a fluorophore; wherein thecomposition is suitable for injection or implantation into a human.

In certain embodiments, the composition is a cohesive gel. Thecomposition may be a hydrated gel. The composition may be transparent ortranslucent.

In certain embodiments, the tissue filler medium retains the fluorophorewithin the composition during administering of the composition, and atleast during a portion of the illumination. Alternatively, thefluorophore may leach from the tissue filler medium after administeringto the tissue.

In certain embodiments, the tissue filler medium is biodegradable. Thetissue filler medium may comprise any biodegradable and biocompatiblematerial such as hyaluronic acid, collagen, poly-L-lactic acid. Completebiodegradation may occur in about 3-18 months. The fluorophore may ormay not affect the biodegradation rate of the tissue filler medium.

In certain embodiments, the tissue filler medium comprises cross-linkedhyaluronic acid. The cross-linked hyaluronic acid may be about 0.1 toabout 2%, about 2 to about 30%, about 2 to about 25%, about 2 to about20%, about 2 to about 15%, about 2 to about 10%, about 2 to about 5%, 4to about 30%, about 4 to about 25%, about 4 to about 20%, about 4 toabout 15%, about 4 to about 10%, about 4 to about 5%, of thecomposition. In certain embodiments, the cross-linked hyaluronic acid isin particulate form. The particles may be hydrated. The particles may becohesive. In certain embodiments, the fluorophore is within thecross-linked hyaluronic acid particle.

In certain embodiments, the composition further comprises an injectablemedium supporting the particles. The injectable medium may be a fluid.The injectable medium may be less viscous and/or less cohesive than theparticles. For example, the particles may be cohesive and the injectablemedium non-cohesive. The fluorophore may be in the injectable medium orthe particles, or both. In certain embodiments, the injectable mediumcomprises hyaluronic acid which is non-cross linked or relatively lesscross-linked than the cross-linked hyaluronic acid particles.

In certain embodiments, the composition further comprises lightreflecting particles. The light reflecting particles may be a glass orsilicon dioxide.

In certain embodiments, the fluorophore is a hydrophilic chromophore.The fluorophore may be water soluble. In certain embodiments, thefluorophore is not in a liposomal form in the composition.

In certain embodiments, the fluorophore can be activated by light havinga wavelength in the visible range. In certain embodiments, thefluorophore does not absorb light in the UV range of the electromagneticspectrum. In certain embodiments, when activated, the fluorophore canemit light having a wavelength in the visible range. The emitted lightcan be within one or more of the violet, blue, green, yellow, orange orred portions of the electromagnetic spectrum. Alternatively, thefluorophore can emit in the infrared range.

In certain embodiments, the fluorophore can be photobleached afterillumination of the area. In certain embodiments, the fluorophore may bephotobleached after 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 or 65minutes of illumination. In certain embodiments, the fluorophore may bephotobleached after 1-30 seconds, 15-45 seconds, 30-60 seconds, 0.75-1.5minutes, 1-2 minutes, 1.5-2.5 minutes, 2-3 minutes, 2.5-3.5 minutes, 3-4minutes, 3.5-4.5 minutes, 4-5 minutes, 5-10 minutes, 10-15 minutes,15-20 minutes, 20-25 minutes, or 25-30 minutes. The treatment time mayrange up to about 90 minutes, about 80 minutes, about 70 minutes, about60 minutes, about 50 minutes, about 40 minutes or about 30 minutes. Theillumination may be continuous or pulsed. The illumination may comprisere-illumination of the composition or the area after a few hours ordays.

In certain embodiments, the fluorophore is not a photosensitizing agentrequiring to be metabolized. For example, in certain embodiments, thefluorophore is not porphyrin, porphyrinogen, hematoporphyrin,pheophorbide, chlorin, bacteriochlorin, iso-bacteriochlorin and dihydro-and tetrahydro-tetrapyrroles. In certain embodiments, the fluorophoredoes not bind to native cells, cancer cells or other cellularstructures.

In certain embodiments the composition does not include glucosamine. Incertain embodiments where the composition comprises hyaluronic acid, thecomposition does not include glucosamine. In certain embodiments, thecomposition does not include a source of oxygen such as peroxide. Incertain embodiments, the composition does not include hydrophobicfluorophores. In certain embodiments, the composition is notphotopolymerizable. In certain embodiments, the composition does notinclude a monomer. In certain embodiments, the composition does notinclude hyaluronic acid with functionalized chains. In certainembodiments of any of the foregoing or following, the composition doesnot include one or more (e.g., 1, 2 or 3) of triethanolamine (TEA),N-vinyl-2-pyrrolidone (NVP), or N-vinyl caprolactam (NVC). In certainembodiments, the composition does not include any of triethanolamine(TEA), N-vinyl-2-pyrrolidone (NVP), or N-vinyl caprolactam (NVC).

In certain embodiments of any of the foregoing or following, thecomposition is a sterile composition. In certain embodiments, thecomposition can be sterilized by heat and/or pressure, such as using anautoclave. In certain embodiments, the composition can be sterilized bygamma irradiation. In certain embodiments of any of the above aspects,the subdermally or intradermally placed composition can provide animmediate ‘lifting’ which may be due in part to its cohesive orviscoelastic properties. This is beneficial in smoothing of lines andfolds on skin to provide an appearance of skin rejuvenation. In certainembodiments, the ‘lifting’ effect of the subdermally or intradermallyplaced composition diminishes over time as the tissue filler mediumdegrades, is compressed and/or is dispersed. Therefore, by means ofcertain embodiments of the present disclosure, a prolonged skinrejuvenation effect can be obtained by providing an immediate liftingeffect to the skin which gradually diminishes with time, and at the sametime synthesis of new collagen at or around the intradermal or subdermalcomposition which increases over time. As the tissue filler degradationand the collagen synthesis dovetail into one another, a substantiallycontinuous skin rejuvenation effect may be achieved.

In certain embodiments, the new collagen on or around the intracorporealcomposition substantially matches the extracellular matrix in terms ofcollagen type, collagen mechanical properties and collagen fibrillarorientation. This is unlike existing degradable tissue filler mediawhich either induce collagen synthesis but do not provide a liftingeffect (e.g. fillers based on poly-L-lactic acid), or which provide alifting effect but which do not induce significant collagen synthesis(e.g. hyaluronic acid based dermal fillers), or which provide a liftingeffect and induce collagen synthesis having different properties to theextracellular matrix (e.g. permanent dermal fillers).

According to various aspects, the present disclosure relates to the useof a composition as described herein, for stimulating collagen synthesiswithin soft tissues. From another aspect, there is provided use of acomposition as described herein, for cosmetic enhancement of softtissues. From another aspect, there is provided use of a composition asdescribed herein, for inhibiting or treating scarring. From anotheraspect, there is provided use of a composition as described herein, forskin rejuvenation. From another aspect, there is provided use of acomposition as described herein, together with a topical biophotoniccomposition for skin rejuvenation.

According to various aspects, the present disclosure relates to the useof a composition comprising a tissue filler medium and a fluorophore ina method for stimulating collagen synthesis, wherein the methodcomprises a step of administration of the composition to an area to betreated within a soft tissue, and a step illumination of the area withlight having a wavelength which can be absorbed by the fluorophore; andwherein the method stimulates collagen synthesis in the area.

According to various aspects, the present disclosure relates to the useof a composition comprising a tissue filler medium and a fluorophore ina method for cosmetic enhancement of soft tissue, wherein the methodcomprises a step of administration of the composition to an area to betreated within a soft tissue, and a step illumination of the area withlight having a wavelength which can be absorbed by the fluorophore; andwherein the method cosmetically enhances the soft tissue.

According to various aspects, the present disclosure relates to the useof a composition comprising a tissue filler medium and a fluorophore ina method for inhibiting or treating scarring, wherein the methodcomprises a step of administration of the composition to an area to betreated within a soft tissue, and a step illumination of the area withlight having a wavelength which can be absorbed by the fluorophore; andwherein the method cosmetically inhibits or treats scarring.

According to various aspects, the present disclosure relates a tissuefiller composition comprising more than one tissue filler medium,wherein the composition is suitable for injection or implantation into ahuman.

According to various aspects, the present disclosure relates a methodfor stimulating collagen synthesis comprising: administering a firstcomposition to an area to be treated within a soft tissue, wherein thefirst composition comprises a tissue filler medium; administering on asurface over the area to be treated a second composition comprising afluorophore; and illuminating the area with light having a wavelengthwhich can be absorbed by the fluorophore; wherein the method stimulatescollagen synthesis in the area.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph illustrating the Stokes' shift.

FIG. 2 is a picture depicting the absorption of light in the variouslayers of the skin (Samson et al. Evidence Report/Technology Assessment2004, 111, pages 1-97).

FIG. 3 is a graph illustrating the absorption and emission spectra ofdonor and acceptor chromophores. The spectral overlap between theabsorption spectrum of the acceptor chromophore and the emissionspectrum of the donor chromophore is also shown.

FIG. 4 is a schematic representation of a Jablonski diagram thatillustrates the coupled transitions involved between a donor emissionand acceptor absorbance.

DETAILED DESCRIPTION

The present disclosure provides compositions which can be used as tissuefillers and which are biophotonic. The tissue filler compositions of thepresent disclosure include a photoactive exogenous chromophore togetherwith a tissue filler medium, and may be injectable or implantable ordeliverable to soft tissues in any other way. The present disclosurealso provides methods useful for stimulating collagen synthesis;cosmetic enhancement of soft tissue such as soft tissue augmentation,rejuvenation or re-shaping; inhibition or treatment of scars; orpromoting wound healing, using such tissue filler compositions.

Before continuing to describe the present disclosure in further detail,it is to be understood that this disclosure is not limited to specificcompositions or process steps, as such may vary.

It must be noted that, as used in this specification and the appendedclaims, the singular form “a”, “an” and “the” include plural referentsunless the context clearly dictates otherwise.

As used herein, the term “about” in the context of a given value orrange refers to a value or range that is within 20%, preferably within10%, and more preferably within 5% of the given value or range.

It is convenient to point out here that “and/or” where used herein is tobe taken as specific disclosure of each of the two specified features orcomponents with or without the other. For example “A and/or B” is to betaken as specific disclosure of each of (i) A, (ii) B and (iii) A and B,just as if each is set out individually herein.

“Biophotonic” means the generation, manipulation, detection andapplication of photons in a biologically relevant context. In otherwords, biophotonic compositions exert their physiological effectsprimarily due to the generation and manipulation of photons. Biophotoniccompositions may also generate reactive oxygen species. “Biophotoniccomposition” is a composition as described herein that may be activatedby light to produce photons and/or reactive oxygen species forbiologically relevant applications.

Terms “chromophore”, “photoactivating agent” and “photoactivator” areused herein interchangeably. A chromophore means a chemical compound,when contacted by light irradiation, is capable of absorbing the light.The chromophore readily undergoes photoexcitation and can then transferits energy to other molecules and/or emit it as light. The chromophoremay be a synthetic chromophore or a naturally occurring chromophore.

“Fluorophore” as used herein means a chromophore which can emit lightupon light excitation e.g. by fluorescence, phosphorescence or any othermeans.

“Tissue filler” when used herein means a material that is suitable for,or generally used for, tissue augmentation, regeneration or re-shapingsuch as those suitable for use or used in the dermis area such as dermalfillers, or those suitable for use or used in any other soft tissue forexample as a scaffold or a delivery material. Tissue fillers includebiodegradable and non-biodegradable materials, as well as natural andsynthetic materials, including hydrogels. Tissue fillers can beadministered by any means such as by injection or implantation. Tissuefillers can be administered to any soft tissue site such assubcutaneous, hypodermic and/or intradermal. “Dermal filler” when usedherein means a material which can be used in the dermis area, such asbelow the epidermis and/or above or below the hypodermis, such as fortissue augmentation, regeneration or re-shaping. Dermal fillers can bedelivered by hypodermic and/or intradermal injection, or by any othermeans such as implantation.

As used herein, the expression “soft tissue” refers to tissues thatconnect, support, or surround other structures and organs of the body,not being bone. Soft tissue includes tendons, ligaments, fascia, skin,fibrous tissues, fat, and synovial membranes (which are connectivetissue), and muscles, nerves and blood vessels (which are not connectivetissue). It is sometimes defined by what it is not. Soft tissue has beendefined as “nonepithelial, extraskeletal mesenchyme exclusive of thereticuloendothelial system and glia”.

“Injectable” when used herein means a flowable material which may bedrawn through or pushed through a needle by a syringe for injection intoa soft tissue. Soft tissue includes tendons, ligaments, fascia, skin,dermis, fibrous tissues, fat, synovial membranes, muscles, nerves andblood vessels.

“Inhibiting or treating scarring” as used herein means preventing orminimizing scar formation, or reducing an existing scar. The scarringcan be a result of any wound such as a burn or a surgical incision.

“Photobleaching” means the photochemical destruction of a chromophore.

The term “light” or “actinic light” is intended to mean light energyemitted from a specific light source (e.g., lamp, LED, laser) andcapable of being absorbed by matter (e.g. the chromophore orphotoactivator defined above). In a preferred embodiment, the light hasa peak wavelength within the visible range of the electromagneticspectrum, e.g. 360 nm to 760 nm.

“Wound” means an injury to any tissue, including for example, acute,subacute, delayed or difficult to heal wounds, and chronic wounds.Examples of wounds may include both open and closed wounds. Woundsinclude, for example, amputations, burns, incisions, excisions, lesions,lacerations, abrasions, puncture or penetrating wounds, surgical wounds,contusions, hematomas, crushing injuries, ulcers (such as for examplepressure, venous, arterial or diabetic), wounds caused by periodontitis(inflammation of the periodontium).

“Skin rejuvenation” means the reducing, diminishing, retarding orreversing of one or more signs of skin aging or skin damage. Skinrejuvenation also means generally improving the cosmetic appearance ofskin or cosmetic enhancement of skin. For instance, it includesincreasing luminosity of the skin; reducing pore size; reducing finelines or wrinkles; improving thin and transparent skin; improvingfirmness; improving plumpness; augmenting tissue loss or sagging skindue to loss of underlying fat or bone; improving dry skin (which mightitch); reducing or reversing freckles, age spots, spider veins, or ablotchy complexion; improving rough and leathery skin; or improving finewrinkles that disappear when stretched. According to the presentdisclosure, one or more of the above conditions may be improved or oneor more of the above signs of aging may be reduced, diminished, retardedor even reversed by certain embodiments of the compositions, methods anduses of the present disclosure.

In some embodiments, the present disclosure provides biophotoniccompositions which can be injected or implanted within soft tissue astissue fillers. The present disclosure provides intracorporealbiophotonic compositions. Biophotonic compositions are compositions thatare, in a broad sense, activated by light (e.g., photons) of specificwavelength. These compositions contain at least one exogenouschromophore which can be activated by light having an appropriatewavelength causing the chromophore to emit light. The activating and/oremitted light may have a therapeutic effect on its own. The activatedchromophore and/or the light may also lead to the photochemicalactivation of other agents contained in the composition or at atreatment site. For example, in the presence of an oxygen-releasingagent, such an agent may be activated leading to the formation of oxygenradicals, such as singlet oxygen. The oxygen-releasing agent may be aninternal source of oxygen such as blood.

In some aspects, the present disclosure provides biophotoniccompositions comprising at least a first chromophore and a tissue fillermedium. When a chromophore absorbs a photon of a certain wavelength, itbecomes excited. This is an unstable condition and the molecule tries toreturn to the ground state, giving away the excess energy. For somechromophores, it is favorable to emit the excess energy as light whentransforming back to the ground state. This process is calledfluorescence and these chromophores are also known as “fluorophores”.The peak wavelength of the emitted fluorescence is shifted towardslonger wavelengths compared to the absorption wavelengths due to loss ofenergy in the conversion process. This is called the Stokes' shift andis illustrated in FIG. 1. In the proper environment (e.g., in abiophotonic composition) much of this energy is transferred to othercomponents of the composition or to the treatment site directly.

Without being bound to theory, it is thought that fluorescent lightemitted by photoactivated chromophores may have therapeutic propertiesdue to its femto-, pico- or nano-second emission properties which may berecognized by biological cells and tissues, leading to favorablebiomodulation. Furthermore, the emitted fluorescent light has a longerwavelength and hence a deeper penetration into the tissue than theactivating light (FIG. 2). Irradiating intradermal, subdermal or othersoft tissue with such a broad range of wavelengths, including in someembodiments the activating light which passes through the composition,may have different and complementary effects on the cells and tissues.

In certain embodiments, the tissues surrounding the biophotoniccomposition will be illuminated with wavelengths of light which wouldnot normally reach those soft tissue sites if illuminated transdermallydue to the short wavelengths and the depth from the skin. In certainembodiments, this may lead to a therapeutic or cosmetic benefit to thesoft tissues surrounding the biophotonic composition in use, such asstimulation of collagen production. This may be in addition to anyphysical filling and lifting effects of a typical tissue filler, as wellas in addition to any collagen production due induced by locally appliedstresses from the tissue filler. Furthermore, the collagen which isproduced may more closely match that of the extracellular collagenmatrix of the area being treated compared to fibrous collagen formationthrough foreign body response seen with some existing tissue fillers.That is the type of collagen, the mechanical properties of the collagen,and collagen fibrillar direction will more closely match that of nativecollagen.

In some instances, the biophotonic compositions of the presentdisclosure may be capable of triggering changes in cell signalling as soto produce an increase in collagen deposition through fibroblaststimulation and thereby modifying the extracellular matrix locally.

The biophotonic compositions of the present disclosure may besubstantially transparent/translucent and/or have high lighttransmittance in order to permit light dissipation into and through thecomposition. In this way, the area of tissue around the composition canbe treated both with the fluorescent light emitted by the compositionand the light irradiating the composition to activate it. The %transmittance of the biophotonic composition can be measured in therange of wavelengths from 250 nm to 800 nm using, for example, aPerkin-Elmer Lambda 9500 series UV-visible spectrophotometer. In someembodiments, transmittance within the visible range is measured andaveraged. In some other embodiments, transmittance of the biophotonicmaterial is measured with the chromophore omitted. In some embodiments,transmittance of the compositions disclosed herein is measured at 460nm. As transmittance is dependent upon thickness, the thickness of eachsample can be measured with calipers prior to loading in thespectrophotometer. Transmittance values can be normalized to a thicknessof 100 μm (or any thickness) according to the following formula:

${{F_{T\text{-}{corr}}\left( {\lambda,t_{2}} \right)} = {\left\lbrack {{e^{- \sigma_{t}}(\lambda)}t_{1}} \right\rbrack^{\frac{t_{2}}{t_{1}}} = \left\lbrack {F_{t\text{-}{corr}}\left( {\lambda,t_{1}} \right)} \right\rbrack^{\frac{t_{2}}{t_{1}}}}},$where t₁=actual specimen thickness, t₂=thickness to which transmittancemeasurements can be normalized. In the art, transmittance measurementsare usually normalized to 1 cm. In some embodiments, the biophotoniccomposition has a transmittance that exceeds about 15%, about 20%, about30%, about 40%, about 50%, about 55%, about 60%, about 65%, about 70%,about 75%, about 80%, or about 85% within the visible range. In someembodiments, the transmittance exceeds about 70%, about 86%, about 87%,about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about94%, about 95%, about 96%, about 97%, about 98% or about 99% within thevisible range of the electromagnetic spectrum (e.g. 400-700 nm).

The biophotonic compositions of the present disclosure are injectable orimplantable, or deliverable intracorporeally to a soft tissue site byany other means.

These compositions may be described based on the components making upthe composition. Additionally or alternatively, the compositions of thepresent disclosure have functional and structural properties and theseproperties may also be used to define and describe the compositions.Individual components of the composition of the present disclosure aredetailed as below.

The compositions of the present disclosure comprise one or morechromophores, which can be considered exogenous, e.g., are not naturallypresent in skin or tissue. Suitable chromophores can be fluorophores orfluorochromes such as fluorescent dyes (or stains), although other dyegroups or dyes (biological and histological dyes, food colorings,carotenoids, naturally occurring fluorescent and other dyes) can also beused. Suitable photoactivators can be those that are Generally RegardedAs Safe (GRAS). Photoactivators which are not well tolerated by the skinor other tissues can be included in the biophotonic composition in anencapsulated, or chemically modified form.

In certain embodiments, the biophotonic composition of the presentdisclosure comprises a first chromophore which undergoes partial orcomplete photobleaching upon application of light. By photobleaching ismeant a photochemical destruction of the chromophore which can generallybe visualized as a loss of color. In some embodiments, the firstchromophore absorbs at a wavelength in the range of the visiblespectrum, such as at a wavelength of about 380-800 nm, about 380-700,about 400-700 or about 380-600 nm. In these embodiments, the firstchromophore is not activated by UV light. In other embodiments, thefirst chromophore absorbs at a wavelength of about 200-800 nm, about200-700 nm, about 200-600 nm or about 200-500 nm. In one embodiment, thefirst chromophore absorbs at a wavelength of about 200-600 nm. In someembodiments, the first chromophore absorbs light at a wavelength ofabout 200-300 nm, about 250-350 nm, about 300-400 nm, about 350-450 nm,about 400-500 nm, about 400-600 nm, about 450-650 nm, about 600-700 nm,about 650-750 nm or about 700-800 nm.

It will be appreciated by those skilled in the art that opticalproperties of a particular chromophore may vary depending on thechromophore's surrounding medium. Therefore, as used herein, aparticular chromophore's absorption and/or emission wavelength (orspectrum) corresponds to the wavelengths (or spectrum) measured in abiophotonic composition of the present disclosure.

The biophotonic compositions disclosed herein may include at least oneadditional chromophore. Combining chromophores may increasephoto-absorption by the combined dye molecules and enhance absorptionand photo-biomodulation selectivity. This creates multiple possibilitiesof generating new photosensitive, and/or selective chromophore mixtures.

When such multi-chromophore compositions are illuminated with light,energy transfer can occur between the chromophores. This process, knownas resonance energy transfer, is a photophysical process through whichan excited ‘donor’ chromophore (also referred to herein as firstchromophore) transfers its excitation energy to an ‘acceptor’chromophore (also referred to herein as second chromophore). Theefficiency and directedness of resonance energy transfer depends on thespectral features of donor and acceptor chromophores. In particular, theflow of energy between chromophores is dependent on a spectral overlapreflecting the relative positioning and shapes of the absorption andemission spectra. For energy transfer to occur, the emission spectrum ofthe donor chromophore must overlap with the absorption spectrum of theacceptor chromophore (FIG. 3). Energy transfer manifests itself throughdecrease or quenching of the donor emission and a reduction of excitedstate lifetime accompanied also by an increase in acceptor emissionintensity. FIG. 4 is a Jablonski diagram that illustrates the coupledtransitions involved between a donor emission and acceptor absorbance.To enhance the energy transfer efficiency, the donor chromophore shouldhave good abilities to absorb photons and emit photons. Furthermore, itis thought that the more overlap there is between the donorchromophores' emission spectra and the acceptor chromophore's absorptionspectra, the better a donor chromophore can transfer energy to theacceptor chromophore.

In certain embodiments, the biophotonic composition of the presentdisclosure further comprises a second chromophore. In some embodiments,the first chromophore has an emission spectrum that overlaps at leastabout 80%, at least about 50%, at least about 40%, at least about 30%,at least about 20%, or at least about 10% with an absorption spectrum ofthe second chromophore. In one embodiment, the first chromophore has anemission spectrum that overlaps at least about 20% with an absorptionspectrum of the second chromophore. In some embodiments, the firstchromophore has an emission spectrum that overlaps at least about 1-10%,at least about 5-15%, at least about 10-20%, at least about 15-25%, atleast about 20-30%, at least about 25-35%, at least about 30-40%, atleast about 35-45%, at least about 50-60%, at least about 55-65% or atleast about 60-70% with an absorption spectrum of the secondchromophore.

% spectral overlap, as used herein, means the % overlap of a donorchromophore's emission wavelength range with an acceptor chromophore'sabsorption wavelength range, measured at spectral full width quartermaximum (FWQM). For example, FIG. 2 shows the normalized absorption andemission spectra of donor and acceptor chromophores. The spectral FWQMof the acceptor chromophore's absorption spectrum is from about 60 nm(515 nm to about 575 nm). The overlap of the donor chromophore'sspectrum with the absorption spectrum of the acceptor chromophore isabout 40 nm (from 515 nm to about 555 nm). Thus, the % overlap can becalculated as 40 nm/60 nm×100=66.6%.

In some embodiments, the second chromophore absorbs at a wavelength inthe range of the visible spectrum. In certain embodiments, the secondchromophore has an absorption wavelength that is relatively longer thanthat of the first chromophore within the range of about 50-250, about25-150 or about 10-100 nm.

As discussed above, the application of light to the compositions of thepresent disclosure can result in a cascade of energy transfer betweenthe chromophores. In certain embodiments, such a cascade of energytransfer provides emission of photons from inside the soft tissue thatcan travel deeper than if the photons were travelling transdermally. Insome embodiments, such a cascade of energy transfer is not accompaniedby concomitant generation of heat. In some other embodiments, thecascade of energy transfer does not result in tissue damage.

Optionally, when the biophotonic composition comprises a first and asecond chromophore, the first chromophore is present in an amount ofabout 0.001-40% per weight of the composition, and the secondchromophore is present in an amount of about 0.001-40% per weight of thecomposition. In certain embodiments, the total weight per weight ofchromophore or combination of chromophores may be in the amount of about0.001-40.001% per weight of the composition. In certain embodiments, thefirst chromophore is present in an amount of about 0.001-0.01%, about0.001-0.05%, about 0.005-0.01%, about 0.01-1%, about 0.01-2%, about0.05-1%, about 0.05-2%, about 1-5%, about 2.5-7.5%, about 5-10%, about7.5-12.5%, about 10-15%, about 12.5-17.5%, about 15-20%, about17.5-22.5%, about 20-25%, about 22.5-27.5%, about 25-30%, about27.5-32.5%, about 30-35%, about 32.5-37.5%, or about 35-40% per weightof the composition. In certain embodiments, the second chromophore ispresent in an amount of about 0.001-1%, about 0.001-2%, about0.001-0.01%, about 0.01-0.1%, about 0.1-1.0%, about 1-2%, about 1-5%,about 2.5-7.5%, about 5-10%, about 7.5-12.5%, about 10-15%, about12.5-17.5%, about 15-20%, about 17.5-22.5%, about 20-25%, about22.5-27.5%, about 25-30%, about 27.5-32.5%, about 30-35%, 3 about2.5-37.5%, or about 35-40% per weight of the composition. In certainembodiments, the total weight % per weight of chromophore or combinationof chromophores may be in the amount of about 0.001-1%, about 0.01-1%,about 0.01-2%, about 0.05-2%, about 0.5-1%, about 0.5-2%, about 1-5%,about 2.5-7.5%, about 5-10%, about 7.5-12.5%, about 10-15%, about12.5-17.5%, about 15-20%, about 17.5-22.5%, about 20-25%, about22.5-27.5%, about 25-30%, about 27.5-32.5%, about 30-35%, about32.5-37.5%, or about 35-40.05% per weight of the composition.

In some embodiments, the chromophore or chromophores are selected suchthat their emitted fluorescent light, on photoactivation, is within oneor more of the blue, green, yellow, orange, red and infrared portions ofthe electromagnetic spectrum, for example having a peak wavelengthwithin the range of about 450 nm to about 500 nm, 490 nm to about 800nm, or about 470 nm to about 700 nm. In certain embodiments, the emittedfluorescent light has a peak power density of between 0.005 to about 10mW/cm², or about 0.5 to about 5 mW/cm².

Suitable chromophores that may be used in the biophotonic compositionsof the present disclosure include, but are not limited to the following:

Chlorophyll dyes—Exemplary chlorophyll dyes include but are not limitedto chlorophyll a; chlorophyll b; oil soluble chlorophyll;bacteriochlorophyll a; bacteriochlorophyll b; bacteriochlorophyll c;bacteriochlorophyll d; protochlorophyll; protochlorophyll a; amphiphilicchlorophyll derivative 1; and amphiphilic chlorophyll derivative 2.

Xanthene derivatives—Exemplary xanthene dyes include but are not limitedto Eosin B (4′,5′-dibromo,2′,7′-dinitro-fluorescein, dianion); eosin Y;eosin Y (2′,4′,5′,7′-tetrabromo-fluorescein, dianion); eosin(2′,4′,5′,7′-tetrabromo-fluorescein, dianion); eosin(2′,4′,5′,7′-tetrabromo-fluorescein, dianion) methyl ester; eosin(2′,4′,5′,7′-tetrabromo-fluorescein, monoanion) p-isopropylbenzyl ester;eosin derivative (2′,7′-dibromo-fluorescein, dianion); eosin derivative(4′,5′-dibromo-fluorescein, dianion); eosin derivative(2′,7′-dichloro-fluorescein, dianion); eosin derivative(4′,5′-dichloro-fluorescein, dianion); eosin derivative(2′,7′-diiodo-fluorescein, dianion); eosin derivative(4′,5′-diiodo-fluorescein, dianion); eosin derivative(tribromo-fluorescein, dianion); eosin derivative(2′,4′,5′,7′-tetrachloro-fluorescein, dianion); eosin; eosindicetylpyridinium chloride ion pair; erythrosin B(2′,4′,5′,7′-tetraiodo-fluorescein, dianion); erythrosin; erythrosindianion; erythiosin B; fluorescein; fluorescein dianion; phloxin B(2′,4′,5′,7′-tetrabromo-3,4,5,6-tetrachloro-fluorescein, dianion);phloxin B (tetrachloro-tetrabromo-fluorescein); phloxine B; rose bengal(3,4,5,6-tetrachloro-2′,4′,5′,7′-tetraiodofluorescein, dianion); pyroninG, pyronin J, pyronin Y; Rhodamine dyes such as rhodamines include4,5-dibromo-rhodamine methyl ester; 4,5-dibromo-rhodamine n-butyl ester;rhodamine 101 methyl ester; rhodamine 123; rhodamine 6 G; rhodamine 6 Ghexyl ester; tetrabromo-rhodamine 123; and tetramethyl-rhodamine ethylester.

Methylene blue dyes—Exemplary methylene blue derivatives include but arenot limited to 1-methyl methylene blue; 1,9-dimethyl methylene blue;methylene blue; methylene blue (16 μM); methylene blue (14 μM);methylene violet; bromomethylene violet; 4-iodomethylene violet;1,9-dimethyl-3-dimethyl-amino-7-diethyl-amino-phenothiazine; and1,9-dimethyl-3-diethylamino-7-dibutyl-amino-phenothiazine.

Azo dyes—Exemplary azo (or diazo-) dyes include but are not limited tomethyl violet, neutral red, para red (pigment red 1), amaranth(Azorubine S), Carmoisine (azorubine, food red 3, acid red 14), allurared AC (FD&C 40), tartrazine (FD&C Yellow 5), orange G (acid orange 10),Ponceau 4R (food red 7), methyl red (acid red 2), and murexide-ammoniumpurpurate.

In some aspects of the disclosure, the one or more chromophores of thebiophotonic composition disclosed herein can be independently selectedfrom any of Acid black 1, Acid blue 22, Acid blue 93, Acid fuchsin, Acidgreen, Acid green 1, Acid green 5, Acid magenta, Acid orange 10, Acidred 26, Acid red 29, Acid red 44, Acid red 51, Acid red 66, Acid red 87,Acid red 91, Acid red 92, Acid red 94, Acid red 101, Acid red 103, Acidroseine, Acid rubin, Acid violet 19, Acid yellow 1, Acid yellow 9, Acidyellow 23, Acid yellow 24, Acid yellow 36, Acid yellow 73, Acid yellowS, Acridine orange, Acriflavine, Alcian blue, Alcian yellow, Alcoholsoluble eosin, Alizarin, Alizarin blue 2RC, Alizarin carmine, Alizarincyanin BBS, Alizarol cyanin R, Alizarin red S, Alizarin purpurin,Aluminon, Amido black 10B, Amidoschwarz, Aniline blue WS, Anthraceneblue SWR, Auramine O, Azocannine B, Azocarmine G, Azoic diazo 5, Azoicdiazo 48, Azure A, Azure B, Azure C, Basic blue 8, Basic blue 9, Basicblue 12, Basic blue 15, Basic blue 17, Basic blue 20, Basic blue 26,Basic brown 1, Basic fuchsin, Basic green 4, Basic orange 14, Basic red2 (Saffranin O), Basic red 5, Basic red 9, Basic violet 2, Basic violet3, Basic violet 4, Basic violet 10, Basic violet 14, Basic yellow 1,Basic yellow 2, Biebrich scarlet, Bismarck brown Y, Brilliant crystalscarlet 6R, Calcium red, Carmine, Carminic acid (acid red 4), Celestineblue B, China blue, Cochineal, Coelestine blue, Chrome violet CG,Chromotrope 2R, Chromoxane cyanin R, Congo corinth, Congo red, Cottonblue, Cotton red, Croceine scarlet, Crocin, Crystal ponceau 6R, Crystalviolet, Dahlia, Diamond green B, DiOC6, Direct blue 14, Direct blue 58,Direct red, Direct red 10, Direct red 28, Direct red 80, Direct yellow7, Eosin B, Eosin Bluish, Eosin, Eosin Y, Eosin yellowish, Eosinol, Eriegarnet B, Eriochrome cyanin R, Erythrosin B, Ethyl eosin, Ethyl green,Ethyl violet, Evans blue, Fast blue B, Fast green FCF, Fast red B, Fastyellow, Fluorescein, Food green 3, Gallein, Gallamine blue, Gallocyanin,Gentian violet, Haematein, Haematine, Haematoxylin, Helio fast rubinBBL, Helvetia blue, Hematein, Hematine, Hematoxylin, Hoffman's violet,Imperial red, Indocyanin green, Ingrain blue, Ingrain blue 1, Ingrainyellow 1, INT, Kermes, Kermesic acid, Kemechtrot, Lac, Laccaic acid,Lauth's violet, Light green, Lissamine green SF, Luxol fast blue,Magenta O Magenta I, Magenta II, Magenta III, Malachite green,Manchester brown, Martius yellow, Merbromin, Mercurochrome, Metanilyellow, Methylene azure A, Methylene azure B, Methylene azure C,Methylene blue, Methyl blue, Methyl green, Methyl violet, Methyl violet2B, Methyl violet 10B, Mordant blue 3, Mordant blue 10, Mordant blue 14,Mordant blue 23, Mordant blue 32, Mordant blue 45, Mordant red 3,Mordant red 11, Mordant violet 25, Mordant violet 39 Naphthol blueblack, Naphthol green B, Naphthol yellow S, Natural black 1, Naturalred, Natural red 3, Natural red 4, Natural red 8, Natural red 16,Natural red 25, Natural red 28, Natural yellow 6, NBT, Neutral red, Newfuchsin, Niagara blue 3B, Night blue, Nile blue, Nile blue A, Nile blueoxazone, Nile blue sulphate, Nile red, Nitro BT, Nitro blue tetrazolium,Nuclear fast red, Oil red O, Orange G, Orcein, Pararosanilin, PhloxineB, phycobilins, Phycocyanins, Phycoerythrins. Phycoerythrincyanin (PEC),Phthalocyanines, Picric acid, Ponceau 2R, Ponceau 6R, Ponceau B, Ponceaude Xylidine, Ponceau S, Primula, Purpurin, Pyronin B, Pyronin G, PyroninY, Rhodamine B, Rosanilin, Rose bengal, Saffron, Safranin O, Scarlet R,Scarlet red, Scharlach R, Shellac, Sirius red F3B, Solochrome cyanin R,Soluble blue, Solvent black 3, Solvent blue 38, Solvent red 23, Solventred 24, Solvent red 27, Solvent red 45, Solvent yellow 94, Spiritsoluble eosin, Sudan III, Sudan IV, Sudan black B, Sulfur yellow S,Swiss blue, Tartrazine, Thioflavine S, Thioflavine T, Thionin, Toluidineblue, Toluyline red, Tropaeolin G, Trypaflavine, Trypan blue, Uranin,Victoria blue 4R, Victoria blue B, Victoria green B, Water blue I, Watersoluble eosin, Xylidine ponceau, or Yellowish eosin.

In certain embodiments, the composition of the present disclosureincludes any of the chromophores listed above, or a combination thereof,so as to provide a biophotonic impact at the treatment site. This is adistinct application of these agents and differs from the use ofchromophores as simple stains or as a catalyst for photo-polymerization.In certain embodiments, the composition does not include compounds whichcan be polymerized or cross-linked through activation of thechromophore.

Without being bound to any particular theory, a synergistic effect ofthe chromophore combinations means that the biophotonic effect isgreater than the sum of their individual effects. Advantageously, thismay translate to increased reactivity of the biophotonic material,faster or improved treatment time. Also, the treatment conditions neednot be altered to achieve the same or better treatment results, such astime of exposure to light, power of light source used, and wavelength oflight used. In other words, use of synergistic combinations ofchromophores may allow the same or better treatment withoutnecessitating a longer time of exposure to a light source, a higherpower light source or a light source with different wavelengths.

In some embodiments, the material includes Eosin Y as a firstchromophore and any one or more of Rose Bengal, Fluorescein,Erythrosine, Phloxine B, chlorophyllin as a second chromophore. It isbelieved that these combinations have a synergistic effect as they cantransfer energy to one another when activated due in part to overlaps orclose proximity of their absorption and emission spectra. Thistransferred energy is then emitted as fluorescence or leads toproduction of reactive oxygen species. This absorbed and re-emittedlight is thought to be transmitted throughout the composition, and alsoto be transmitted into the site of treatment.

In further embodiments, the material includes the following synergisticcombinations: Eosin Y and Fluorescein; Fluorescein and Rose Bengal;Erythrosine in combination with Eosin Y, Rose Bengal or Fluorescein;Phloxine B in combination with one or more of Eosin Y, Rose Bengal,Fluorescein and Erythrosine. Other synergistic chromophore combinationsare also possible.

By means of synergistic effects of the chromophore combinations in thematerial, chromophores which cannot normally be activated by anactivating light (such as a blue light from an LED), can be activatedthrough energy transfer from chromophores which are activated by theactivating light. In this way, the different properties ofphotoactivated chromophores can be harnessed and tailored according tothe cosmetic or the medical therapy required.

For example, Rose Bengal can generate a high yield of singlet oxygenwhen activated in the presence of molecular oxygen; however it has a lowquantum yield in terms of emitted fluorescent light. Rose Bengal has apeak absorption around 540 nm and so can be activated by green light.Eosin Y has a high quantum yield and can be activated by blue light. Bycombining Rose Bengal with Eosin Y, one obtains a composition which canemit therapeutic fluorescent light and generate singlet oxygen whenactivated by blue light. In this case, the blue light photoactivatesEosin Y which transfers some of its energy to Rose Bengal as well asemitting some energy as fluorescence.

The present disclosure provides biophotonic compositions that compriseat least a first chromophore and a tissue filler medium. The tissuefiller medium may be a dermal filler. In some embodiments, the tissuefiller medium is characterized by its source. In some embodiments, thesource can be natural, biologic or synthetic. Biologic tissue fillermedia can be those that are derived from a living organism. In someembodiments, the tissue filler medium can be characterized by the body'sability to clear a product without external intervention (e.g.,biodegradable vs. non-biodegradable). The tissue filler media and/or thebiophotonic composition is generally biocompatible. The tissue fillermedia and/or the biophotonic composition is generally non-toxic. Thetissue filler medium may comprise a polymer. The polymer may be selectedfrom the group of polymers consisting of proteins, peptides,polypeptides, polylysine, collagens, pro-collagens, elastins, andlaminins. The polymer may be selected from the group of polymersconsisting of synthetic polymers with hydroxyl, amine, and carboxylfunctional groups: poly(vinyl alcohol), polyethylene glycol, polyvinlylamine, polyallylamine, deacetylated polyacrylamide, polyacrylic acid,and polymethacrylic acid. The polymer may be selected from the group ofpolymers consisting of dendritic or branched polymers, includingdendritic polyols and dendritic polyamines. The polymer may be selectedfrom the group of polymers consisting of solid surface with hydroxyl,amine, and carboxyl functional groups. The polymer may be apolysaccharide, for example, selected from the group of polysaccharidesincluding starch and its derivatives; dextran and its derivatives,cellulose and its derivatives; chitin and chitosan and alginate and itsderivatives. In some embodiments, the tissue filler medium is across-linked biocompatible polysaccharide gel.

In some embodiments, the polymer is a glycosaminoglycan. The tissuefiller medium can further comprise two or more differentglycosaminoglycan polymers. As used herein, the term “glycosaminoglycan”is synonymous with “GAG” and “mucopolysaccharide” and refers to longunbranched polysaccharides consisting of a repeating disaccharide units.The repeating unit consists of a hexose (six-carbon sugar) or ahexuronic acid, linked to a hexosamine (six-carbon sugar containingnitrogen) and pharmaceutically acceptable salts thereof. Members of theGAG family vary in the type of hexosamine, hexose or hexuronic acid unitthey contain, such as, e.g., glucuronic acid, iduronic acid, galactose,galactosamine, glucosamine) and may also vary in the geometry of theglycosidic linkage. Non-limiting examples of glycosaminoglycans includechondroitin sulfate, dermatan sulfate, keratan sulfate, and hyaluronan.Non-limiting examples of an acceptable salt of a glycosaminoglycansincludes sodium salts, potassium salts, magnesium salts, calcium salts,and combinations thereof. Glycosaminoglycan and their resulting polymersuseful in the compositions and methods disclosed herein are describedin, e.g., Piron and Tholin, Polysaccharide Crosslinking, HydrogelPreparation, Resulting Polysaccharides(s) and Hydrogel(s), uses Thereof,U.S. Patent Publication 2003/0148995; Lebreton, Cross-Linking of Low andHigh Molecular Weight Polysaccharides Preparation of InjectableMonophase Hydrogels; Lebreton, Viscoelastic Solutions Containing SodiumHyaluronate and Hydroxypropyl Methyl Cellulose, Preparation and Uses,U.S. Patent Publication 2008/0089918; Lebreton, Hyaluronic Acid-BasedGels Including Lidocaine, U.S. Patent Publication 2010/0028438; andPolysaccharides and Hydrogels thus Obtained, U.S. Patent Publication2006/0194758; and Di Napoli, Composition and Method for Intradermal SoftTissue Augmentation, international Patent Publication WO 2004/073759,each of which is hereby incorporated by reference in its entirety. GAGsuseful in the methods disclosed herein are commercially available, suchas, e.g., hyaluronan-based dermal fillers JUVEDERM™, JUVEDERM™ 30,JUVEDERM™ Ultra, JUVEDERM™ Ultra Plus, JUVEDERM™ Ultra XC, JUVEDERM™Ultra Plus XC, JUVEDERM VOLUMA™ XC, and JUVEDERM VOLUMA™ (Allergan Inc.,Irvine, Calif.).

Examples of biologic, biodegradable tissue filler media are those thatinclude materials derived from organism, human, and/or animal tissuesand/or products. Examples of such media include the following:hyaluronic acid (HA), (such as the following: avian HA, bovine HA, andnon-animal stabilized HA (“NASHA”) (e.g., RESTYLANE®, Captique® andJuvederm® injectable dermal fillers)), collagen (such as collagen I,collagen II, collagen III, cross-linked and/or non-cross-linked, bovine,porcine, human, and autologous collagen). Additional examples ofcollagen based fillers include ZYPLAST® (collagen derived from bovinetissue), ZYDERM® I (collagen derived from bovine tissue), ZYDERM® II,(collagen derived from bovine tissue), EVOLENCE™ (porcine derivedcollagen), and FIBREL™ (porcine derived collagen). Collagen-based tissuefillers are generally animal derived and have been associated with ahigher occurrence of allergic reactions than non-animal based fillers.NASHA tissue fillers, for example, are bacteria derived and have a lowerincidence of allergic reaction than the collagen-based fillers. ManyNASHA tissue fillers provide an immediate filling effect but can alsoinduce minimal collagen formation due to local mechanical deformation offibroblasts at the soft tissue site. As can be appreciated by one ofskill in the art, in some embodiments, the filler media isself-replicating, and can include living cells (such ascollagen-producing cells or fibroblasts). Thus, in some embodiments thecomposition comprises a tissue filler medium that is biologic andbiodegradable.

Synthetic, biodegradable, tissue filler media include RADIANCE™ andRADIESSE™ (calcium and phosphate based microspheres), LARESSE®(carboxymethyl cellulose and polyethylene oxide), SCULPTRA®(microspheres of poly-L-lactic acid), other polyacids, polyethers andpolymers. The calcium and phosphate based microspheres comprise calciumhydroxylapatite particles suspended in a water-based gel that acts as ascaffold for new collagen growth. The particles degrade over time intocalcium and phosphate ions which can be removed by normal metabolicprocesses. Degradation typically takes up to 18 months or even longer.Fillers based on poly-L-lactic acid are considered as “stimulatory” asthey are thought to stimulate collagen production over time. Therefore,an immediate filling effect is not seen. Also, they have been associatedwith granuloma and nodule formation.

Synthetic, non-biodegradable, tissue filler media include compounds thatare not readily broken down in the body. Synthetic, non-biodegradable,tissue filler media can include a biologic component (and vice versa).In some embodiments, at least a portion of the product cannot besignificantly broken down by various body processes. Examples ofsynthetic non-biodegradable filler media include the following: ARTEFIL™and ARTECOL™ (polymethylmethacrylate (PMMA) microspheres suspended inbovine collagen carrier), SILSKIN® (liquid medical grade silicone),DERMALIVE™ and DERMADEEP™ (stabilized hyaluronic acid plus acrylichydrogel hydroxyethylmethacrylate (HEMA) and ethylmethacrylate (EMA)co-polymer particles) and various other polymers, polyacids, andpolyethers. In some embodiments, the carrier has rapid biodegradation.Some of these non-degradable fillers will stimulate a fibroblasticdeposition of collagen around the non-degradable material as part of thebody's immune response. In some cases, the newly formed collagen may notbe closely matched in type, fibrillar alignment and/or mechanicalproperties to the body's native collagen matrix. Also, complications mayarise due to the non-degradable nature of the material and may be longerlasting and more difficult to treat than with degradable fillers.Furthermore, permanent fillers have also been associated with severefibrotic reactions leading in some cases to scarring.

As can be appreciated by one of skill in the art, in some embodiments,any one, or combination, of ingredients of the above fillers can becombined with the other fillers (or alternative fillers) in variousembodiments and for particular results.

In some embodiments, the tissue filler media is selected from thefollowing: collagen, fat, human or animal derived collagen, bovinecollagen, type I collagen, type II collagen, type III collagen, 3.5%bovine dermal collagen cross-linked by glutaraldehyde to form alatticework, natural human collagen, autologous collagen,polymethylmethacrylate microspheres (optionally suspended in bovinecollagen), suspension of collagen fibers prepared from the subject'stissue, human tissue collagen matrix derived from cadaveric dermis,polyacids and polyethers (e.g., carboxymethyl cellulose (CMC) andpolyethylene oxide), acellular human cadaveric dermis that has beenfreeze-dried, micronized acellular human cadaveric dermis that has beenfreeze-dried, cultured autologous fibroblasts, hyaluronic acid,non-animal-stabilized hyaluronic acid derivative, microspheres ofcalcium hydroxyl apatite suspended in an aqueous gel carrier, dextranbeads suspended in hylan gel of nonanimal origin (e.g., 40- to 60-μm indiameter), solubilized elastin peptides with bovine collagen, silicone,solubilized elastin peptides with bovine collagen, poly-L-lactic acid,polytetrafluoroethylene (PTFE), glycosylated collagen, PMMA,bone-forming calcium apatite, cultured human cells, expanded PTFE(e-PTFE), SOFTFORM® of ePTFE, and some combination thereof. Furtherexamples of injectable filler media include the following: AQUAMID®(comprising water and cross-linked polymers), ARTEFIL® (PMMAmicrospheres suspended in bovine collagen), LARESSE® Dermal Filler(synthetic, biocompatible polymers, non-HA gel comprising absorbablemedical polymers), ARTECOLL® (PMMA microspheres suspended in bovinecollagen), BELOTERO®, BIO-ALCAMID™ (synthetic reticulate polymer(poly-alkyl-imide), CAPTIQUE™ (non-animal hyaluronic acid), COSMODERM™(human collagen skin filler), COMOPLAST™, CYMETRA®, autologen,DERMALOGEN®, FASCIAN™ (fascia), fascia, fat, Hylaform™ (avian hyaluronicacid), JUVEDERM®, JUVEDERM VOLBELLA, JUVEDERM VOLUMA, JUVADERM ESTHELIS,JUVADERM FORTELIS, RESTYLANE®, PERLANE® (biosynthesized, non-animalhyaluronic acid), RADIESSE™ (microspheres based on calcium andphosphate), SCULPTRA® (poly-L-lactic acid (PLLA)), collagen, hyaluronicacid, ZYDERM®, ZYPLAST® (collagen derived from bovine tissue),DERMALIVE®, (hyaluronic acid and acrylic hydrogel particles), DERMADEEP®(hyaluronic acid and acrylic hydrogel particles), HYDRAFILL®, ISOLAGEN®(cultured autologous human fibroblasts), LARESSE®(carboxymethylcellulose (CMC) and polyethylene oxide (PEO) filler),PURAGEN™ (filler comprising double cross-linked hyaluron molecules),REVIDERM® INTRA (filler comprising flexible dextran micro-beadssuspended in super-coiled, stabilized hyaluronic acid), SCULPTRA™(Formerly NEW-FILL™, filler from poly-L-lactic acid), Teosyal,SURGIDERM® (hyaluronic acid filler involving 3D hyaluronic acid matrixtechnology), OUTLINE®, ANIKA®, Cosmetic tissue augmentation (CTA, fromAnika), and combinations thereof.

Other examples of tissue fillers include: Juvederm® VOLIFT® fromAllergan, Emervel® from Galderm, Elevess® from Anika Therapeutics,Regenovue® from Neogenesis.

In some embodiments, additional agents may be combined with the tissuefiller medium. The agent combined with the polymer may comprise ananaesthetizing agent or a painkilling agent (e.g. lidocaine), or avitamin (e.g. vitamin C). The additional agent may comprise abiologically active component such as growth factors, peptides andcells.

In some embodiments, the tissue filler medium is substantially stable atroom temperature for, e.g., about 3 months, about 6 months, about 9months, about 12 months, about 15 months, about 18 months, about 21months, about 24 months, about 27 months, about 30 months, about 33months, or about 36 months. In other aspects of this embodiment, adermal filler composition is substantially stable at room temperaturefor, e.g., at least 3 months, at least 6 months, at least 9 months, atleast 12 months, at least 15 months, at least 18 months, at least 21months, at least 24 months, at least 27 months, at least 30 months, atleast 33 months, or at least 36 months. In other aspects of thisembodiment, a tissue filler composition is substantially stable at roomtemperature for, e.g., about 3 months to about 12 months, about 3 monthsto about 18 months, about 3 months to about 24 months, about 3 months toabout 30 months, about 3 months to about 36 months, about 6 months toabout 12 months, about 6 months to about 18 months, about 6 months toabout 24 months, about 6 months to about 30 months, about 6 months toabout 36 months, about 9 months to about 12 months, about 9 months toabout 18 months, about 9 months to about 24 months, about 9 months toabout 30 months, about 9 months to about 36 months, about 12 months toabout 18 months, about 12 months to about 24 months, about 12 months toabout 30 months, about 12 months to about 36 months, about 18 months toabout 24 months, about 18 months to about 30 months, or about 18 monthsto about 36 months.

In some embodiments, the tissue filler medium is degradable and degradesin vivo in about 1 month, 2 months, 3 months, about 3 months to about 12months, about 3 months to about 18 months, about 3 months to about 24months, about 6 months to about 12 months, about 6 months to about 18months, about 9 months to about 12 months, about 9 months to about 18months, about 12 months to about 18 months.

In certain embodiments, the tissue filler medium comprises cross-linkedhyaluronic acid. The hyaluronic acid may be non-animal derived such asthrough a streptococcal fermentation process which is then stabilized bycross-linking. Such bacterially derived hyaluronic acids have a shorterchain length and molecular weight than animal based hyaluronic acid(about 1-3 megadaltons compared with 4-6 megadaltons).

Cross-linking of hyaluronic acid can be achieved using cross-linkingagents such as 1, 4-butanediol diglycidyl ether (BDDE) (e.g. Restylane,Juvederm and Boletero dermal fillers), divinyl sulphone (DVS) (e.g.Hylaform, Captique and Prevelle dermal fillers), 2, 7, 8-diepoxyoctane(DEO), a polyethylene glycol based crosslinking agent as described inU.S. Patent Application Publication No: 2014/0039062, the contents ofwhich are incorporated herein by reference, and biscarbodiimide in thepresence of a pH buffer, as described in U.S. Pat. No. 8,124,120, thecontents of which are incorporated herein by reference. Cross-linkingrenders the hyaluronic acid slower to degrade. The type and extent ofcross-linking will determine the rate of degradation, viscoelasticproperties as well as the stability of the tissue filler medium.However, a possible limitation on the extent of cross-linking may be adecrease in biocompatibility (e.g. increased inflammatory response andgranuloma formation in vivo) and a high viscosity rendering the fillernon-injectable.

The tissue filler medium may comprise a relatively insolublecross-linked hyaluronic acid component within a soluble fluid component.The cross-linked hyaluronic acid component may comprise cohesiveparticles. The soluble fluid component may comprise free hyaluronic acid(non cross-linked) or any other fluid component which can facilitate thedelivery of the tissue filler composition through fine bore needles. Theconcentration of hyaluronic acid (including both cross-linked and noncross-linked hyaluronic acid components) can vary from about 4.5 toabout 40 mg/mL, about 4.5 to about 35 mg/mL, about 4.5 to about 30mg/mL, about 4.5 to about 25 mg/mL, about 4.5 to about 20 mg/mL, about4.5 to about 15 mg/mL, about 4.5 to about 10 mg/mL. The concentration ofhyaluronic acid (including both cross-linked and non cross-linkedhyaluronic acid components) can be about 10 mg/mL, about 12 mg/mL, about14 mg/mL, about 16 mg/mL, about 18 mg/mL, about 20 mg/mL, about 22mg/mL, or about 24 mg/mL. The non cross-linked hyaluronic acid componentmay comprise from about 30% to about 100% of the total hyaluronic acid,or about 40-100%, about 40-99%, about 40-98%, about 40-95%, about40-90%, about 40-85%, about 40-80%, about 40-75%, about 40-70%, about40-65%, about 40-60%, about 40-55%, about 40-50%, about 40-45%, about50-100%, about 50-99%, about 50-98%, about 50-95%, about 50-90%, about50-85%, about 50-80%, about 50-75%, about 50-70%, about 50-65%, about50-60%, about 50-55%, about 60-100%, about 60-99%, about 60-98%, about60-95%, about 60-90%, about 60-85%, about 60-80%, about 60-75%, about60-70% of the total hyaluronic acid content in the composition. Thecross-linked hyaluronic acid component may comprise from about 1% toabout 20% of the total hyaluronic acid content in the composition. Incertain embodiments, the cross-linked hyaluronic acid componentcomprises from about 1% to about 15%, about 1% to about 14%, about 1% toabout 13%, about 1% to about 12%, about 1% to about 11%, about 1% toabout 10%, about 1% to about 9%, about 1% to about 8%, about 1% to about7%, about 1% to about 6%, about 1% to about 5%, about 1% to about 4%,about 1% to about 3%, about 1% to about 2%. In certain embodiments, thecross-linked hyaluronic acid component comprises from about 1% to about12%, about 3% to about 10%, or about 4% to about 10%.

In certain embodiments, the tissue filler medium comprises cross-linkedhyaluronic acid particles within a fluid component, the particles may bethe same or different sizes. The particles may be sized to enable themto pass through a fine needle bore such as a 27-40G needle. The particlesizes may range from 100-1000 microns, or about 200-1000 microns, about250-1000 microns, about 300-1000 microns, about 350-1000 microns, about400-1000 microns about 450-1000 microns, about 500-1000 microns, about550-1000 microns, about 600-1000 microns, about 650-1000 microns, about700-1000 microns, about 200-900 microns, about 250-900 microns, about300-900 microns, about 350-900 microns, about 400-900 microns about450-900 microns, about 500-900 microns, about 550-900 microns, about600-900 microns, about 650-900 microns, about 700-900 microns, 200-800microns, about 250-800 microns, about 300-800 microns, about 350-800microns, about 400-800 microns about 450-800 microns, about 500-800microns, about 550-800 microns, about 600-800 microns, about 650-800microns, or about 700-800 microns.

Cohesiveness of the tissue filler composition may be a desired propertyin applications where it is desired to mechanically lift soft tissuearound the implantation or injection site. The elastic modulus G′ isoften used to characterize the firmness of a gel, and represents thematerial's ability to resist deformation. In hyaluronic gels, the degreeof cross-linking and hyaluronic acid concentration affects the modulusof the composition. Increasing hyaluronic acid concentration andcross-linking will increase the filler modulus. In certain embodiments,the elastic modulus, G′, of the tissue filler media ranges from 100-800Pa, about 100-700 Pa, about 100-600 Pa, about 100-500 Pa, about 200-600Pa, or about 200-500 Pa.

In certain embodiments, the tissue filler medium is hydrated. The extentof hydration is a factor in determining how much the medium will swellonce administered to soft tissue.

The biophotonic compositions of the present disclosure have numeroususes. Without being bound by theory, the biophotonic compositions of thepresent disclosure may be used for skin rejuvenation. The biophotoniccompositions of the present disclosure may promote wound healing ortissue repair. The biophotonic compositions of the present disclosuremay provide cosmetic enhancement of soft tissue. The biophotoniccompositions of the present disclosure may inhibit or treat scarring.The biophotonic compositions of the present disclosure may stimulatecollagen synthesis. This collagen synthesis may be useful for tissuerepair, skin rejuvenation, or cosmetic enhancement of soft tissue.Therefore, it is an objective of the present disclosure to provide amethod for providing biophotonic therapy to a wound, where the methodpromotes wound healing. It is an objective of the present disclosure toprovide a method for providing biophotonic therapy to a wound, where themethod promotes collagen synthesis. It is an objective of the presentdisclosure to provide a method for providing biophotonic therapy to awound, where the method prevents scar formation. It is also an objectiveof the present disclosure to provide a method for providing biophotonictherapy to skin tissue, wherein the method is used for promoting skinrejuvenation. It is also an objective of the present disclosure toprovide a method for providing biophotonic therapy to skin tissue,wherein the method is used for promoting collagen synthesis.

In certain embodiments, the present disclosure provides a method forproviding a biophotonic therapy to a wound, the method comprising:administering a composition to an area to be treated within a wound,wherein the composition comprises a tissue filler medium and afluorophore; illuminating the area with light having a wavelength whichcan be absorbed by the fluorophore; wherein the method stimulates woundhealing in the area.

In yet another aspect, the present disclosure provides a method forpromoting skin rejuvenation. In certain embodiments, the presentdisclosure provides a method for providing skin rejuvenation, the methodcomprising: administering a composition to an area to be treated withina soft tissue, wherein the composition comprises a tissue filler mediumand a fluorophore; illuminating the area with light having a wavelengthwhich can be absorbed by the fluorophore; wherein the method stimulatescollagen synthesis in the area. The tissue filler medium may provide animmediate filling effect and the composition may induce collagensynthesis.

In yet another aspect, the present disclosure provides a method forcosmetic enhancement of soft tissue. In certain embodiments, the presentdisclosure provides a method for cosmetic enhancement of soft tissuecomprising: intradermally or subdermally administering a composition toan area to be treated, wherein the composition comprises a tissue fillermedium and a fluorophore; illuminating the area with light having awavelength which can be absorbed by the fluorophore; wherein the methodstimulates collagen synthesis in the area to cosmetically enhance thesoft tissue.

In yet another aspect, the present disclosure provides a method forinhibiting or treating scarring. In certain embodiments, the presentdisclosure provides a method for inhibiting or treating scarring, themethod comprising: administering a composition to an area to be treatedwithin or around a scar or a wound, wherein the composition comprises atissue filler medium and a fluorophore; illuminating the area with lighthaving a wavelength which can be absorbed by the fluorophore; whereinthe method stimulates collagen synthesis in the area to prevent orreduce scar formation.

In the methods disclosed herein, the biophotonic composition can beadministered by injection or implantation (e.g., using a syringe andneedle, etc.) into or underneath soft tissue at a treatment site (e.g.,subcutaneous administration, intradermal administration, subdermaladministration). A skilled person can select an appropriate needle boresize according to the soft tissue to be treated. In intradermalapplications, needle gauges of 27 G to 40 G can be used, typically 30 or32 G. The higher the gauge size, the finer the needle bore. Thebiophotonic compositions of the disclosure can also be injected orimplanted superficially, such as, for example, within the papillarylayer of the dermis, or can be injected or implanted within thereticular layer of the dermis.

The biophotonic composition can be administered intradermally orsubdermally into the dermis in a continuous fashion (e.g. linearthreading) or using pin pricks to form discrete pockets of thebiophotonic composition intradermally or subdermally (e.g. serialpuncture technique). The biophotonic composition can be illuminated atthe same time as administering the composition to the soft tissue site,or after administration.

In certain embodiments, about 1 mL of the biophotonic composition isadministered intradermally or subdermally. In certain embodiments, lessthan about 1 mL of the biophotonic composition is administeredintradermally or subdermally. In certain embodiments, about 0.1-0.2,about 0.2-0.3, about 0.3-0.4, about 0.4-0.5, about 0.5-0.6, about0.6-0.7, about 0.7-0.8, about 0.8-0.9, about 0.9-1.0 mL of thebiophotonic composition is administered intradermally or subdermally.

The biophotonic compositions suitable for use in the methods of thepresent disclosure may be selected from any of the embodiments of thebiophotonic compositions described above. For instance, the biophotoniccompositions useful in the method of the present disclosure may comprisea first chromophore that undergoes at least partial photobleaching uponapplication of light. The first chromophore may absorb at a wavelengthof about 200-800 nm, about 200-700 nm, about 200-600 nm or about 200-500nm. In one embodiment, the first chromophore absorbs at a wavelength ofabout 200-600 nm. In some embodiments, the first chromophore absorbslight at a wavelength of about 200-300 nm, about 250-350 nm, about300-400 nm, about 350-450 nm, about 400-500 nm, about 450-650 nm, about600-700 nm, about 650-750 nm or about 700-800 nm. In other examples,suitable biophotonic compositions for the methods of the presentdisclosure may further comprise at least one additional chromophore(e.g., a second chromophore). The absorption spectrum of the secondchromophore overlaps at least about 80%, at least about 50%, at leastabout 40%, at least about 30%, or at least about 20% with the emissionspectrum of the first chromophore. In some embodiments, the firstchromophore has an emission spectrum that overlaps at least about 1-10%,at least about 5-15%, at least about 10-20%, at least about 15-25%, atleast about 20-30%, at least about 25-35%, at least about 30-40%, atleast about 35-45%, at least about 50-60%, at least about 55-65% or atleast about 60-70% with an absorption spectrum of the secondchromophore.

Illumination of the biophotonic composition with light may cause atransfer of energy from the first chromophore to the second chromophore.Subsequently, the second chromophore may emit energy as fluorescenceand/or generate reactive oxygen species. In certain embodiments of themethods of the present disclosure, energy transfer caused by theapplication of light is not accompanied by concomitant generation ofheat, or does not result in tissue damage.

In the methods of the present disclosure, the biophotonic compositionmay be illuminated transdermally (from outside the body) or from withinthe soft tissue (e.g. using an optical fibre or a light duct). Incertain embodiments, the biophotonic composition may itself form a lightduct from the skin surface to the composition. To form such a lightduct, a trail of the composition is formed from the skin puncture pointto the composition within the soft tissue. In the methods of the presentdisclosure, the biophotonic composition may be illuminated at the sametime as or immediately following administration of the composition tothe intracorporeal area.

In the methods of the present disclosure, any source of actinic lightcan be used. Any type of halogen, LED or plasma arc lamp or laser may besuitable. The primary characteristic of suitable sources of actiniclight will be that they emit light in a wavelength (or wavelengths)appropriate for activating the one or more photoactivators present inthe composition. In one embodiment, an argon laser is used. In anotherembodiment, a potassium-titanyl phosphate (KTP) laser (e.g. aGreenLight™ laser) is used. In another embodiment, sunlight may be used.In yet another embodiment, a LED photocuring device is the source of theactinic light. In yet another embodiment, the source of the actiniclight is a source of light having a wavelength between about 200 to 800nm. In another embodiment, the source of the actinic light is a sourceof visible light having a wavelength between about 400 and 600 nm orabout 400 to 700 nm. In yet another embodiment, the source of theactinic light is blue light. In yet another embodiment, the source ofthe actinic light is red light. In yet another embodiment, the source ofthe actinic light is green light. Furthermore, the source of actiniclight should have a suitable power density. Suitable power density fornon-collimated light sources (LED, halogen or plasma lamps) are in therange from about 1 mW/cm² to about 200 mW/cm².

Suitable power density for laser light sources are in the range fromabout 0.5 mW/cm² to about 0.8 mW/cm².

In some embodiments of the methods of the present disclosure, the lighthas an energy at the subject's skin, wound or mucosa surface of betweenabout 1 mW/cm² and about 500 mW/cm², 1-300 mW/cm², or 1-200 mW/cm²,wherein the energy applied depends at least on the condition beingtreated, the wavelength of the light, the distance of the subject's skinfrom the light source, and the thickness of the biophotonic composition.In certain embodiments, the light at the subject's skin is between about1-40 mW/cm², or 20-60 mW/cm², or 40-80 mW/cm², or 60-100 mW/cm², or80-120 mW/cm², or 100-140 mW/cm², or 120-160 mW/cm², or 140-180 mW/cm²,or 160-200 mW/cm², or 110-240 mW/cm², or 110-150 mW/cm², or 190-240mW/cm².

In some embodiments of the methods of the present disclosure, abiophotonic topical composition may be and additional source ofillumination or the only source of illumination. In these embodiments,the biophotonic topical composition may comprise a fluorophore such thatwhen illuminated with an activating light (e.g. from an LED or laserlight source) it will emit light having a longer wavelength (Stoke'sshift). The light from the activating light and/or the biophotonictopical composition may activate the fluorophore within the tissuefiller composition. In this way, the tissues surrounding the tissuefiller composition are illuminated with a broad bandwidth of light ofdifferent intensities. The topical biophotonic compositions may be asdescribed in any one of WO 2010/051636, WO 2010/051641 and WO2013/155620, the contents of which are herein incorporated by reference.

In certain embodiments, the chromophore(s) in the composition can bephotoexcited by ambient light including from the sun and overheadlighting. In certain embodiments, the chromophore(s) can bephotoactivated by light in the visible range of the electromagneticspectrum. The light can be emitted by any light source such as sunlight,light bulb, an LED device, electronic display screens such as on atelevision, computer, telephone, mobile device, flashlights on mobiledevices. In the methods of the present disclosure, any source of lightcan be used. For example, a combination of ambient light and directsunlight or direct artificial light may be used. Ambient light caninclude overhead lighting such as LED bulbs, fluorescent bulbs etc. andindirect sunlight.

The duration of the exposure to actinic light required will be dependenton depth beneath the skin surface of the biophotonic composition; thethickness, density and components of the intervening tissue; the type ofintervening tissue, the concentration of chromophore within the tissuefiller composition, the power density, wavelength and bandwidth of thelight source, the thickness of the biophotonic composition, and thetreatment distance from the light source. The illumination of thetreated area by fluorescence may take place within seconds or evenfragments of seconds, but a prolonged exposure period is beneficial toexploit the synergistic effects of the absorbed, reflected and reemittedlight on the composition of the present disclosure and its interactionwith the tissue being treated.

In one embodiment, the time of exposure to actinic light of the tissuein which the biophotonic composition has been administered is a periodbetween 1 minute and 5 minutes. In another embodiment, the time ofexposure to actinic light of the tissue in which the biophotoniccomposition has been administered is a period between 1 minute and 5minutes. In some other embodiments, the biophotonic composition isilluminated for a period between 1 minute and 3 minutes. In certainembodiments, light is applied for a period of 1-30 seconds, 15-45seconds, 30-60 seconds, 0.75-1.5 minutes, 1-2 minutes, 1.5-2.5 minutes,2-3 minutes, 2.5-3.5 minutes, 3-4 minutes, 3.5-4.5 minutes, 4-5 minutes,5-10 minutes, 10-15 minutes, 15-20 minutes, 20-25 minutes, or 20-30minutes. The treatment time may range up to about 90 minutes, about 80minutes, about 70 minutes, about 60 minutes, about 50 minutes, about 40minutes or about 30 minutes. It will be appreciated that the treatmenttime can be adjusted in order to maintain a dosage by adjusting the rateof fluence delivered to a treatment area. For example, the deliveredfluence may be about 4 to about 60 J/cm², about 10 to about 60 J/cm²,about 10 to about 50 J/cm², about 10 to about 40 J/cm², about 10 toabout 30 J/cm², about 20 to about 40 J/cm², about 15 J/cm² to 25 J/cm²,or about 10 to about 20 J/cm².

In yet another embodiment, the source of actinic light is in continuousmotion over the treated area for the appropriate time of exposure. Inyet another embodiment, multiple applications of the biophotoniccomposition and actinic light are performed. In some embodiments, thebiophotonic composition or the tissue is exposed to actinic light atleast two, three, four, five or six times. In some embodiments, a freshapplication of the topical biophotonic composition is applied beforeexposure to actinic light, or a fresh biophotonic compositionadministered to the soft tissues.

The method may be repeated as necessary. For example, in certainembodiments where the tissue filler medium is a biodegradable material,the method may be repeated close to or after full degradation of thetissue filler medium. The tissue filler medium degradation time dependson the type of filler medium and its inherent degradation propertiessuch as viscosity, the quantity of filler in the soft tissues, itsplacement within the soft tissues, and the depth of the tissue. Theperiod of repeating the method according to certain embodiments of thepresent disclosure can range from about 2 months up to about 24 months.In certain embodiments, the fluorophore is not degraded afterillumination, in which case the composition can be re-illuminated untilphotobleaching.

The method may further comprise massaging the area once the compositionhas been administered.

The epidermis and the dermis are the first and second layers of skin,respectively. The dermis contains the structural elements of the skin,the connective tissue. There are various types of connective tissue withdifferent functions. Elastin fibers give the skin its elasticity, andcollagen gives the skin its strength.

The junction between the dermis and the epidermis is an importantstructure. The dermal-epidermal junction interlocks forming finger-likeepidermal ridges. The cells of the epidermis receive their nutrientsfrom the blood vessels in the dermis. The epidermal ridges increase thesurface area of the epidermis that is exposed to these blood vessels andthe needed nutrients.

Aging of skin comes with significant physiological changes to the skin.The generation of new skin cells slows down, and the epidermal ridges ofthe dermal-epidermal junction flatten out. While the number of elastinfibers increases, their structure and coherence decrease. Also theamount of collagen and the thickness of the dermis decrease with theageing of the skin.

Collagen is a major component of the skin's extracellular matrix,providing a structural framework. During the aging process, the decreaseof collagen synthesis and insolubilization of collagen fibers contributeto a thinning of the dermis and loss of the skin's biomechanicalproperties.

The physiological changes to the skin result in noticeable agingsymptoms often referred to as chronological-, intrinsic- andphoto-ageing. The skin becomes drier, roughness and scaling increase,the appearance becomes duller, and most obviously fine lines andwrinkles appear.

Other symptoms or signs of skin aging include, but are not limited to,thinning and transparent skin, loss of underlying fat (leading tohollowed cheeks and eye sockets as well as noticeable loss of firmnesson the hands and neck), bone loss (such that bones shrink away from theskin due to bone loss, which causes sagging skin), dry skin (which mightitch), inability to sweat sufficiently to cool the skin, unwanted facialhair, freckles, age spots, spider veins, rough and leathery skin, finewrinkles that disappear when stretched, loose skin, a blotchycomplexion.

The dermal-epidermal junction is a basement membrane that separates thekeratinocytes in the epidermis from the extracellular matrix, which liesbelow in the dermis. This membrane consists of two layers: the basallamina in contact with the keratinocytes, and the underlying reticularlamina in contact with the extracellular matrix. The basal lamina isrich in collagen type IV and laminin, molecules that play a role inproviding a structural network and bioadhesive properties for cellattachment.

Laminin is a glycoprotein that only exists in basement membranes. It iscomposed of three polypeptide chains (alpha, beta and gamma) arranged inthe shape of an asymmetric cross and held together by disulfide bonds.The three chains exist as different subtypes which result in twelvedifferent isoforms for laminin, including Laminin-1 and Laminin-5.

The dermis is anchored to hemidesmosomes, specific junction pointslocated on the keratinocytes, which consist of α-integrins and otherproteins, at the basal membrane keratinocytes by type VII collagenfibrils. Laminins, and particularly Laminin-5, constitute the realanchor point between hemidesmosomal transmembrane proteins in basalkeratinocytes and type VII collagen.

Laminin-5 synthesis and type VII collagen expression have been proven todecrease in aged skin. This causes a loss of contact between dermis andepidermis, and results in the skin losing elasticity and becoming saggy.

Another type of wrinkles, generally referred to as expression wrinkles,require loss of resilience, particularly in the dermis, because of whichthe skin is no longer able to resume its original state when facialmuscles which produce facial expressions exert stress on the skin,resulting in expression wrinkles.

The compositions and methods of the present disclosure promote skinrejuvenation. In certain embodiments, the compositions and methods ofthe present disclosure promote collagen synthesis. In certain otherembodiments, the compositions and methods of the present disclosure mayreduce, diminish, retard or even reverse one or more signs of skin agingincluding, but not limited to, appearance of fine lines or wrinkles,thin and transparent skin, loss of underlying fat (leading to hollowedcheeks and eye sockets as well as noticeable loss of firmness on thehands and neck), bone loss (such that bones shrink away from the skindue to bone loss, which causes sagging skin), dry skin (which mightitch), inability to sweat sufficiently to cool the skin, unwanted facialhair, freckles, age spots, spider veins, rough and leathery skin, finewrinkles that disappear when stretched, loose skin, or a blotchycomplexion. In certain embodiments, the compositions and methods of thepresent disclosure may induce a reduction in pore size, enhancesculpturing of skin subsections, and/or enhance skin translucence.Furthermore, the compositions and methods of the present disclosure mayenhance the cosmetic appearance of skin such as by improving luminosityand texture, reducing pore size, and tightening the skin.

The biophotonic materials and methods of the present disclosure may beused to treat wounds, promote wound healing, promote tissue repairand/or prevent or reduce cosmesis including improvement of motorfunction (e.g. movement of joints). Wounds that may be treated by thebiophotonic materials and methods of the present disclosure include, forexample, injuries to the skin and subcutaneous tissue initiated indifferent ways (e.g., pressure ulcers from extended bed rest, woundsinduced by trauma or surgery, burns, ulcers linked to diabetes or venousinsufficiency, wounds induced by conditions such as periodontitis) andwith varying characteristics. In certain embodiments, the presentdisclosure provides biophotonic materials and methods for treatingand/or promoting the healing of, for example, burns, incisions,excisions, lesions, lacerations, abrasions, puncture or penetratingwounds, surgical wounds, contusions, hematomas, crushing injuries,amputations, sores and ulcers.

Biophotonic compositions and methods of the present disclosure may beused to treat and/or promote the healing of chronic cutaneous ulcers orwounds, which are wounds that have failed to proceed through an orderlyand timely series of events to produce a durable structural, functional,and cosmetic closure. The vast majority of chronic wounds can beclassified into three categories based on their etiology: pressureulcers, neuropathic (diabetic foot) ulcers and vascular (venous orarterial) ulcers.

For example, the present disclosure provides biophotonic compositionsand methods for treating and/or promoting healing of a diabetic ulcer.Diabetic patients are prone to foot and other ulcerations due to bothneurologic and vascular complications. Peripheral neuropathy can causealtered or complete loss of sensation in the foot and/or leg. Diabeticpatients with advanced neuropathy lose all ability for sharp-dulldiscrimination. Any cuts or trauma to the foot may go completelyunnoticed for days or weeks in a patient with neuropathy. A patient withadvanced neuropathy loses the ability to sense a sustained pressureinsult, as a result, tissue ischemia and necrosis may occur leading tofor example, plantar ulcerations. Microvascular disease is one of thesignificant complications for diabetics which may also lead toulcerations. In certain embodiments, biophotonic materials and methodsof treating a chronic wound are provided here in, where the chronicwound is characterized by diabetic foot ulcers and/or ulcerations due toneurologic and/or vascular complications of diabetes.

In other examples, the present disclosure provides biophotoniccompositions and methods for treating and/or promoting healing of apressure ulcer. Pressure ulcers include bed sores, decubitus ulcers andischial tuberosity ulcers and can cause considerable pain and discomfortto a patient. A pressure ulcer can occur as a result of a prolongedpressure applied to the skin. Thus, pressure can be exerted on the skinof a patient due to the weight or mass of an individual. A pressureulcer can develop when blood supply to an area of the skin is obstructedor cut off for more than two or three hours. The affected skin area canturn red, become painful and necrotic. If untreated, the skin can breakopen and become infected. A pressure ulcer is therefore a skin ulcerthat occurs in an area of the skin that is under pressure from e.g.lying in bed, sitting in a wheelchair, and/or wearing a cast for aprolonged period of time. Pressure ulcers can occur when a person isbedridden, unconscious, unable to sense pain, or immobile. Pressureulcers often occur in boney prominences of the body such as the buttocksarea (on the sacrum or iliac crest), or on the heels of foot.

Additional types of wounds that can be treated by the biophotonicmaterials and methods of the present disclosure include those disclosedby US Patent Application Publication No. 20090220450, which isincorporated herein by reference.

There are three distinct phases in the wound healing process. First, inthe inflammatory phase, which typically occurs from the moment a woundoccurs until the first two to five days, platelets aggregate to depositgranules, promoting the deposit of fibrin and stimulating the release ofgrowth factors. Leukocytes migrate to the wound site and begin to digestand transport debris away from the wound. During this inflammatoryphase, monocytes are also converted to macrophages, which release growthfactors for stimulating angiogenesis and the production of fibroblasts.

Second, in the proliferative phase, which typically occurs from two daysto three weeks, granulation tissue forms, and epithelialization andcontraction begin. Fibroblasts, which are key cell types in this phase,proliferate and synthesize collagen to fill the wound and provide astrong matrix on which epithelial cells grow. As fibroblasts producecollagen, vascularization extends from nearby vessels, resulting ingranulation tissue. Granulation tissue typically grows from the base ofthe wound. Epithelialization involves the migration of epithelial cellsfrom the wound surfaces to seal the wound. Epithelial cells are drivenby the need to contact cells of like type and are guided by a network offibrin strands that function as a grid over which these cells migrate.Contractile cells called myofibroblasts appear in wounds, and aid inwound closure. These cells exhibit collagen synthesis and contractility,and are common in granulating wounds.

Third, in the remodeling phase, the final phase of wound healing whichcan take place from three weeks up to several years, collagen in thescar undergoes repeated degradation and re-synthesis. During this phase,the tensile strength of the newly formed skin increases.

However, as the rate of wound healing increases, there is often anassociated increase in scar formation. Scarring is a consequence of thehealing process in most adult animal and human tissues. Scar tissue isnot identical to the tissue which it replaces, as it is usually ofinferior functional quality. The types of scars include, but are notlimited to, atrophic, hypertrophic and keloidal scars, as well as scarcontractures. Atrophic scars are flat and depressed below thesurrounding skin as a valley or hole. Hypertrophic scars are elevatedscars that remain within the boundaries of the original lesion, andoften contain excessive collagen arranged in an abnormal pattern.Keloidal scars are elevated scars that spread beyond the margins of theoriginal wound and invade the surrounding normal skin in a way that issite specific, and often contain whorls of collagen arranged in anabnormal fashion.

In contrast, normal skin consists of collagen fibers arranged in abasket-weave pattern, which contributes to both the strength andelasticity of the dermis. Thus, to achieve a smoother wound healingprocess, an approach is needed that not only stimulates collagenproduction, but also does so in a way that reduces scar formation.

The biophotonic compositions and methods of the present disclosurepromote wound healing by promoting the formation of substantiallyuniform epithelialization; promoting collagen synthesis; promotingcontrolled contraction; and/or by reducing the formation of scar tissue.In certain embodiments, the biophotonic compositions and methods of thepresent disclosure may promote wound healing by promoting the formationof substantially uniform epithelialization. In some embodiments, thebiophotonic compositions and methods of the present disclosure promotecollagen synthesis. In some other embodiments, the biophotoniccompositions and methods of the present disclosure promote controlledcontraction. In certain embodiments, the biophotonic compositions andmethods of the present disclosure promote wound healing, for example, byreducing the formation of scar tissue. In certain embodiments, thebiophotonic composition can be used during or following wound closure tooptimize scar revision.

The biophotonic composition may be administered at regular intervalssuch as once a week, or at an interval deemed appropriate by thephysician. Alternatively, once administered, the biophotonic compositionmay be light activated at regular intervals until exhaustion of thechromophore.

Adjunct therapies which may be topical or systemic such as antibiotictreatment may also be used. Negative pressure assisted wound closure canalso be used to assist wound closure and/or to remove the composition.

In some instances, the frequency of administration of the biophotoniccompositions as defined herein may depend on the type and/or thequantity of the filler that is initially administered. Fillers can bere-administered as early as a few months and as late as 2 yearsdepending on their thickness, viscosity and elasticity. As such, thequestion of administrations per unit time may depend on the type offiller, the placement, the depth of tissue-site, and the quantityadministered.

The present disclosure also provides kits for preparing and/oradministering any of the compositions of the present disclosure. The kitmay include a container comprising the biophotonic composition of thepresent disclosure. The container may be light impermeable, air-tightand/or leak resistant. Exemplary containers include, but are not limitedto, syringes, vials, or pouches. In some embodiments, the tissue fillermedium and the chromophore compositions are provided in separatecontainers, to be mixed by the user prior to administration. In someembodiments, the tissue filler medium and the chromophore compositionsare provided in a single pre-mixed composition. In some embodiments, thekit comprises a handheld injection device.

In other embodiments, the kit comprises a systemic or topical drug foraugmenting the treatment of the composition. For example, the kit mayinclude a systemic or topical antibiotic or hormone treatment for acnetreatment or wound healing.

Written instructions on how to use the biophotonic composition inaccordance with the present disclosure may be included in the kit, ormay be included on or associated with the containers comprising thecompositions of the present disclosure.

In certain embodiments of the kit, the kit may further comprise a lightsource such as a portable light with a wavelength appropriate toactivate the chromophore in the biophotonic composition. The portablelight may be battery operated or re-chargeable.

In certain embodiments, the kit may further comprise one or morewaveguides.

In certain embodiments, the kit may further comprise topical biophotoniccompositions as described in any one of WO 2010/051636, WO 2010/051641and WO 2013/155620, the contents of which are herein incorporated byreference.

Identification of equivalent compositions, methods and kits are wellwithin the skill of the ordinary practitioner and would require no morethan routine experimentation, in light of the teachings of the presentdisclosure. Practice of the disclosure will be still more fullyunderstood from the following examples, which are presented herein forillustration only and should not be construed as limiting the disclosurein any way.

EXAMPLES

The examples below are given so as to illustrate the practice of thisinvention. They are not intended to limit or define the entire scope ofthis invention.

Example 1

Compositions according to the present disclosure comprising afluorophore and dermal filler were prepared. The following combinationsof dermal filler and fluorophore were used:

-   -   1) JUVEDERM VOLBELLA+Fluorescein+Eosin Y;    -   2) JUVEDERM VOLBELLA+Eosin Y+Eosin B;    -   3) JUVEDERM VOLBELLA+Eosin Y;    -   4) JUVEDERM VOLUMA+Fluorescein+Eosin Y;    -   5) JUVEDERM VOLUMA+Eosin Y;    -   6) JUVEDERM VOLUMA+Eosin Y+Eosin B;    -   7) ESTHELIS+Eosin Y+Fluorescein;    -   8) FORTELIS+Eosin Y+Fluorescein;    -   9) PERLANE®+Fluorescein+Eosin Y;    -   10) PERLANE®+Eosin Y+Eosin B;    -   11) PERLANE®+Eosin Y;    -   12) RESTYLANE®+Fluorescein+Eosin Y;    -   13) RESTYLANE®+Eosin Y+Eosin B; and    -   14) RESTYLANE®+Eosin Y.

For example, compositions 7) and 8) were prepared as follows:

-   -   7) 0.75 g FORTELIS as mixed with 9.38 μl Eosin Y (9.6 mg/ml) and        9.38 μl Fluorescein (9.6 mg/ml) so that the final concentration        of Eosin Y is 0.012% and the final concentration of fluorescein        is 0.012%.    -   8) 0.75 g ESTHELIS was mixed with 9.38 μl Eosin Y (9.6 mg/ml)        and 9.38 μl Fluorescein (9.6 mg/ml) so that the final        concentration of Eosin Y is 0.012% and the final concentration        of fluorescein is 0.012%.

Example 2

Dermal filler-mediated effect on collagen type I and III gene expressionlevel was assessed in Dermal Human Fibroblasts (DHF). Dermal filler gelswere evaluated in vitro to assess their potential effect on collagensynthesis. Collagen is an essential component of the skin extracellularmatrix (ECM) and an increase of collagen synthesis is favorable in skinrejuvenation.

JUVEDERM VOLUMA (VB), JUVADERM VOLBELLA (V15), RESTYLANE® (RL), ESTHELIS(EB) and a mixture of JUVADERM VOLBELLA (V15), RESTYLANE® (RL) andESTHELIS (EB) (Gel A) were used as dermal filler gels in the study. Theeffect of dermal fillers on collagen synthesis was tested in dermalhuman fibroblasts (DHF). Collagen type I and type III mRNA werequantified by qRT-PCR. DHF were cultured on glass bottom dish. Thedermal filler gels were applied on the other side of the glass dish (2mm thick) and were illuminated for 5 min (300 sec) at 5 cm distanceusing blue visible light (KLOX THERA™ lamp). The tested dermal fillergels contained Eosin Y only or the combination of both, Eosin Y andFluorescein (0.011% of each). Dermal filler gels without chromophorewere used as controls.

DHF cells were also treated with light alone without the gel to assessthe effect of the blue light illumination on collagen expressionpattern. Sixteen hours post-illumination, cells were collected for RNAextraction and cDNA synthesis was performed. Collagen mRNA level underdifferent conditions was assessed by qRT-PCR. TGF β1 (5 ng/ml) was usedto trigger collagen genes expression and served as a positive control inall experiments. Results are presented as fold mRNA expression levelcompared to untreated control.

The data is summarized in the Table 1.

TABLE 1 Collagen type I and III mRNA expression in DHF 16 hourspost-treatment compared to untreated control. Type I collagen Type IIIcollagen Dermal filler gels mRNA mRNA Light (KLOX Thera ™ lamp) 1.621.55 VB + no chromophore 0.46 1.1 VB + Eosin Y 2.48 1.16 V15 + nochromophore 0.47 1.58 V15 + Eosin Y 1.7 1.2 V15 + Eosin Y + Fluorescein0.74 1.7 RL + no chromophore 1.17 0.75 RL + Eosin Y 1 0.8 RL + Eosin Y +Fluorescein 1.6 0.98 EB + no chromophore 1.29 1.46 EB + Eosin Y +Fluorescein 1.13 1.64 Gel A + no chromophore 2.3 1.57 Gel A + Eosin Y +Fluorescein 2.9 2.33

Gel A+Eosin Y+Fluorescein showed a positive effect on collagen type Iand type III gene expression (up to 2.9 and 2.33 fold increase,respectively) as compared to untreated control. Interestingly, slightinduction of the collagen type I and type III genes expression was alsoobserved for Gel A no chromophore tested sample (2.3 and 1.57respectively), which could be attributed to the higher ability of Gel Ato transmit blue light which exerts its effect on DHF cells leading tocollagen mRNA expression.

A mixture of tissue filler medium comprising EB, RL and V15 (Gel A)stimulated collagen type I mRNA expression in DHF cells to a higherlevel than the tissue filler medium applied individually as well as incomparison to light alone. Stimulation of collagen type I and collagentype III mRNA expression in DHF cells was increased when chromophoreswere added to Gel A, when compared to Gel A in absence of chromophores.

Example 3

The technique is performed under sterile conditions. The skin in theupper lip area is cleansed with chlorhexidine 0.5% (or similarantiseptic). At that time, a slow dermal or subdermal injection with theHA filler is undertaken. Care is taken not to overfill each area.Following completion of the injection, the area is gently massaged toachieve the desired result. The chromophore gel is applied and the areais illuminated using a LED light for a period of 5 minutes per region ofthe upper lip. Following the treatment with the light, the gel isremoved with a saline wipe.

Example 4

A composition according to the present disclosure comprising afluorophore and a dermal filler comprising hyaluronic acid may beinjected intradermally under a fold on a patient's face. The compositionmay be illuminated with light overlapping the absorption spectra of thefluorophore to photoactivate the fluorophore in situ. The fluorophoremay emit fluorescent light to the surrounding soft tissues.

Example 5

A composition according to the present disclosure comprising afluorophore and a dermal filler comprising hyaluronic acid may beinjected intradermally under a fold on a patient's face. A thin layer ofa topical biophotonic composition may be placed over the fold,immediately above the injected composition. The topical composition maybe illuminated with light overlapping the absorption spectra of afluorophore in the topical composition to photoactivate the fluorophoreand may cause it to emit light having emission spectra which may overlapthe absorption spectra of the fluorophore within the injectedcomposition of the present disclosure. The fluorophore may emitfluorescent light to the surrounding soft tissues.

While the invention has been described in connection with specificembodiments thereof, it will be understood that it is capable of furthermodifications and this application is intended to cover any variations,uses, or adaptations of the invention following, in general, theprinciples of the invention and including such departures from thepresent disclosure as come within known or customary practice within theart to which the invention pertains and as may be applied to theessential features hereinbefore set forth, and as follows in the scopeof the appended claims.

All documents mentioned in the specification are herein incorporated byreference.

The invention claimed is:
 1. A method for promoting wound healing, themethod comprising: administering a composition to a wound area to betreated, wherein the composition comprises a tissue filler medium and afluorophore; and wherein the composition is non-photopolymerizable; andilluminating the wound area to be treated with light having a wavelengthwhich can be absorbed by the fluorophore; wherein the method promoteswound healing in the wound area.
 2. The method of claim 1, wherein thewound area to be treated is soft tissue.
 3. The method of claim 1,wherein the administering of the composition is by injection.
 4. Themethod of claim 1, wherein the administering of the composition is byimplantation.
 5. The method of claim 1, wherein the composition isselected in a form selected from a cohesive gel and a hydrated gel. 6.The method of claim 1, wherein the tissue filler is a dermal filler. 7.The method of claim 1, wherein the tissue filler medium comprises apolymer.
 8. The method of claim 7, wherein the polymer is selected fromproteins, peptides, polypeptides, polylysine, collagens, pro-collagens,elastins, and laminins.
 9. The method of claim 7, wherein the polymer isselected from poly(vinyl alcohol), polyethylene glycol, polyvinlylamine, polyallylamine, deacetylated polyacrylamide, polyacrylic acid,and polymethacrylic acid.
 10. The method of claim 1, wherein the tissuefiller medium comprises cross-linked hyaluronic acid.
 11. The method ofclaim 10, wherein the cross-linked hyaluronic acid is in particulateform.
 12. The method of claim 1, wherein the composition furthercomprises light reflecting particles.
 13. The method of claim 1, whereinthe composition further comprises an injectable medium supporting theparticles.
 14. The method of claim 13, wherein the injectable mediumcomprises hyaluronic acid which is relatively less cross-linked than thehyaluronic acid in particulate form.
 15. The method of claim 1, whereinthe composition is transparent or translucent.
 16. The method of claim1, wherein the tissue filler medium retains the fluorophore within thecomposition during administering of the composition, and at least duringa portion of the illumination.
 17. The method of claim 1, wherein thetissue filler medium is biodegradable.
 18. The method of claim 1,wherein the fluorophore can be activated by light having a wavelength inthe visible range.
 19. The method of claim 1, wherein the fluorophore isnot in a liposomal form in the composition.
 20. The method of claim 1,wherein the wound is an amputation, a burn, an incision, an excision, alesion, a laceration, an abrasion, a puncture wound, a penetratingwound, a surgical wound, a contusion, a hematoma, a crushing injury, apressure ulcer, a venous ulcer, an arterial ulcer, a diabetic ulcer, ora wound caused by periodontitis.